Elements of Applied Microscopy. A Text-Book For Beginners (1905) by Winslow

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ELEMENTS
OF
APPLIED MICROSCOPY.
A TEXT-BOOK FOR BEGINNERS.
CHARLES-EDWARD AMORY WINSLOW,

Instructor in Industrial Microscopy
and Sanitary
Biology in the
Massachusetts Institute of Technology.
FIRST EDITION.
FIRST THOUSAND.
NEW YORK
JOHN WILEY & SONS. London: CHAPMAN & HALL, Limited.

1905.
Copyright, igos,
'
BY
CHARLES-EDWARD AMORY WINSLOW.
ROBERT DRUMMOND, PRINTER, NEW YORK,
'Elements of Applied- Microscopy.
'K
Text-,,,
book for Beginiiers.
By Charles-Edwaed
Amory Winslow,
Instructor in Industrial
Microscopy and Sanitary Biology in the Massachusetts Institute of Technology. Pp. 183, with 60 text figures. New York, John Wiley and Sons. 1905. This manual is an excellent example of a book prepared for a definite purpose and as
result of experience in an institution where independent work and special ideas have a prominent place. As the author states in his preface the book 'Moes not profess to compete on the one hand with monographs or on the other with the popular works on microscopy. It is, however,
,
the
specifically intended for the class in industrial microscopy for second year students in chem-
istry
and biology
at the
Massachusetts Instiobject of the course
tute of Technology.
IS to
The
give facility in the manipulation of the

and. an
microscope
scope of
its
acquaintance
with
thei
practical application.
The
first
four chapters consider the micro-
^scope and
''i3hapters
its accessories,
and the other eight

fibers,
deal with the starches, adulterations
ft food and drugs, textile

fcine
paper, medi-
and
sanitation,
forensic
microscopy,
inicrochemistry, petrology and metallurgy.
Each manner
to
subject
is
dealt
with in a general
to give the student the principles
and
the point of view.

illustrate
Exercises are then given
the
methods necessary for the
elucidation of the questions which arise, in

actual practise.
The book
torily ally
is well conceived and satisfacThe statements are usuworked out. As planned by the clear and concise.
it
author,
is
an introduction
to
the subject,
and was designed
for use by a teacher pos-
DEDICATED BY THE AUTHOR Zo 1bl6 ôtber.
INTRODUCTION.
This
little
book
is
intended for the teacher, and the

specialist.
beginner with the microscope, not for the
It
contains very few original data and treats no single subject with completeness.
Almost
all
the branches of technical microscopy have
been already made the basis for monographs with which

the present volume
can in no
way compete.
as a
On
the
other hand, there are
many
entertaining popular books

it
on the microscope which

study
treat
mere adjunct
to the
of natural history.
Neither type of work was
suited for the use of a class in Industrial Microscopy
which
at
is
offered to second-year Chemists
and
Biologists
the
Massachusetts
is
Institute
of
first,
Technology.
The
object of this course
twofold
to give facility in to fur-
the manipulation of the microscope;
and second,
its
nish an acquaintance with the scope of

application.
practical
brief
As a text-book
treatise
there
was needed a
and elementary
which should take up the fundaitself
mentals of the science and art of microscopy
and
make
a rapid but wide survey of the principal fields in

to
which the microscope has been applied
practical
VI
INTRODUCTION.
affairs.
No
such elementary but comprehensive work

treatises
exists in
Enghsh among the numerous able
upon
is
special branches of the subject.
The
present volume
therefore the outgrowth of a pedagogic need.

is
The book
of
necessarily incomplete
from the standpoint

it
the
if
it
expert in any of the
branches which
treats,
but
conveys to the student's mind such an idea of the possible

applications of the
microscope in varied
fields
as shall
of
stimulate
him
eventually to the further exploration

it
some one
of them,
will
have served
this
its
purpose.
authorities
In the preparation of
volume,
the
quoted in connection with the various chapters have been

freely
drawn upon.
The author
further desires to ex-
press his grateful obligation, to Dr. P. G. Stiles for the
preparation of original drawings;
and
for advice
and
assistance in regard to various portions of the manuscript

to Professor
Professor H.
fessor
W. T. Sedgwick, Professor Mary A. Willcox," M. Goodwin, Professor F. J. Moore, ProWarren, Dr. C. C. Simmons, Mr. A. E.
G.
C. H.
Leach, Mr. A.
Woodman,
Dr.
E. L. Walker, and
Miss A. F. Rogers.
Acknowledgments are due

lishers of the following
to the authors
for figures
and pub-
books
which have
either
been copied directly or redrawn
Deschanel, a.
Elementary Treatise p., and Everett, J. D. on Natural Philosophy. New York, Appleton & Co., 1894. Hager, H., and Mez, C. Das Mikroskop und seine Anwendung-.
Berlin,
J.
Springer, 1899.
Carpenter, W.
B., and Dallinger, W. H. The Microscope and its Revelations. London, J. & A. Churchill, 1891. Gage, S. H. The Microscope. Ithaca., Comstock Publishing
Co., 1904.
INTRODUCTION.
vii
Bausch, E. Use and Care of the Microscope. Rochester, Bausch & Lomb Optical Co., 1902. SCHIMPER, A. F. W. Anleitung zur mikroskopischen Untersuchung der vegetabilischen Nahrungs- und Genusmittel.
Jena, G. Fischer, 1900.
Hassack,
C.
Wodurch
unterscheiden sich die Textilfasern

Berlin, B.
Leipzig-Gohlis, a.
Klepzig, 1900.
J.
Herzberg, W. Papierpriifung. Ultzman, R., und HOFMANN, K.
Springer, 1902.
Atlas der physiologischen
und pathologischen Harnsedimente. Wien, Braumiiller, 1 872. Slater, C, and Spitta, E. J. An Atlas of Bacteriology. London and Philadelphia, J. B. Lippincott Co., 1898. Wesbrook, F. F. Report of the Minnesota State Board of
Health, 1899-1900.
Whipple, G.
C.
The Microscopy
of Drinking-water.
New
York, John Wiley
Galton,
F.
Sons, 1899. Finger-print Directories.
&
London, Macmillan
&
Co., 1895.
Howell, W. H.
Frazer,
1901.
p.
Philadelphia and London,

Bibliotics.
An American W. B.
Text-book of Physiology. Saunders & Co., 1901.

J.
Philadelphia,
B.
Lippincott Co.,
Lehmann,
1891.
O,
Die Krystallanalyse.
Leipzig,
W. Engelmann,
Clark, C. H. Practical Methods in Microscopy. Boston, D. C. Heath & Co., 1894. Luquer, L. M. Minerals in Rock Sections. New York, D.
Sauveur, a.
Van Nostrand Co., 1898. The Constitution
of Steel considered as an Alloy

p, 78.
of Iron and Carbon.
Technology Quarterly, 1898,
CONTENTS.
PAGE
^
I.
Ftinction aot) Parts of the Microscope
II.
Manipulation of the Microscope
23
III.
The Mounting and Preparation

THE Microscope
of Objects for 38
52
IV.
Micrometry, and the Camera Lucida
V.
VI.
VII.
VIII.
The Microscopy
of the Common Starches
58
Foods and Drugs and their Adulterants
69
The Examination The Microscopy The Microscope
of Textile Fibres
80
95
of Paper
in Medicine
IX.
and Sanitation
104
126
141
X.
XI.
XII.
Forensic Microscopy
Microchemistry
Petrography and Metallography
153
LIST OF ILLUSTRATIONS.
PIG.
1.
PAGE
Section through the
Human Eye
2.
Illustration of Refraction
4
5
3. 4.
5.
Refraction by Prisms
Refractions in Prisms of Different Angles
Plano-convex Lens and

to the Principal
its
Principal Focus
to the Relation of a
6.
Course of Rays according
Luminous Point
7
Focus
7.
8.
9.
Formation of Image by Object outside Principal Focus " " " " " " " inside
8 9 ii
12 13
Leeuwenhoek's Microscope
Hooke's Compound Microscope Course of Rays in Compound Microscope
10.
1 1.
12.
13.
Chromatic Aberration
14
15
Achromatic Objective.
14. Effect of 15. 16.
Homogeneous Immersion The Microscope

of
16
18
Relation of Distance from Principal Focus to Size and Position
Image
21
17.
Proper Position for Observer
18.
19.
20.
Method oPInserting Objectives Abbe Condenser Refraction by Air-bubbles and Oil-drops

Micrometer Calipers
24 26 28
,
30
32
33
;
21. Disturbing Effect of the Cover-glass. 22.
23. Curvature of the Field
24. Turntable 25.

26.
27.
35 42
Thoma Microtome
Camera Lucida Course of Rays
in the
48
Camera Lucida
55 56
xn
FIG.
LIST
OF ILLUSTR/ITIONS.
PAGE
,,
28.
The Common
Starches
65 72
75
29. Microscopic Structure of Coffee

30. Microscopic Structure of 31.
Mustard
"
"
"Pepper
77
32. Cotton-fibre 33. Flax-fibre...,. 34.
83 85
Hemp, fibre
Ramie
86
87
35. Jute-fibre 36.
88
91
37. Wool-fibre
38. Silk... 39. Tracheid of a Conifer

40.
93
100
"
" Birch
Id
102
108
41. Cells of Straw
Urinary Sediment in Catarrh of the Bladder " " Acute Bright' s Disease " 4342.
no
1
44. Blood-cells 45. Malarial Parasite

46. Diphtheria Bacilli
13
115
118 122
47. Trichina
48.
Micro-organisms of Drinking-water
124
131
49. Microspectroscope 50. Spectra of

51.
Oxyhsemoglobin and Reduced Haemoglobin
Finger-print Patterns
134 136
139
52.
Minute Characteristics of Handwriting Lines
53. Crystals of 54.

55.
Hydrous and Anhydrous Ferrous Chloride
144
145
145
Crystals of Caesium
Alum
Silver and lodin
Crystals of Calcium Sulphate
Zone between Chlorides of 57. Diagram of a Nicol Prism

56. Contact
58. Interference Figures 59. 60.
148
156
l6i
Curve of
Solidification for Alloy of Silver
and Copper
164
167
Microscopic Appearance of Steel
ELEMENTS OF APPLIED
MICROSCOPY,
CHAPTER
I.
FUNCTION AND PARTS OF THE MICROSCOPE.

I.
Normal Vision.
În
the ordinary act of vision at least
two
distinct processes
may
be distinguished, the formation
upon
seen,
their
the retina (K, Fig. i) of a minute image of the object
and the transformation, by the

end organs,
which we
optic nerves
and
of the energy of the light-waves thus
thrown upon the

activity
retinal surface into that

call sensation.
form
first
of nervous
fol-
The
process
is
lows the simple laws of Optics, while the second

trolled
con-
by the
far
more complex chemical and physical

sci-
conditions which furnish the subject-matter for the
ence of Physiology.
rectly related to the
is,
The
sensation experienced
retina;
is
di-
image lying upon the
that
corre-
to
an area
of special illumination
whose parts
spond
to those of
is
some object
outside,
from which the

learned,
illumination
derived.
The mind has
by
experience, to interpret a certain
image upon the retina
ELEMENTS OF APPLIED MICROSCOPY.

by the
as corresponding to a definite object perceived
other senses.
The image
is
in all cases inverted, but in
infancy
it
is
found that images upside down correspond

really right side up.
to objects
which are
Experience
Fig.
I.
Section through the
Human Eye,
(After Everett-Deschanel.)
A. Cornea. B. Aqueous humor.

C. Pupil.
K. Retina.
M.
N.
D.
Iris.
E. Lens. F. Suspensory ligament. G. Accommodation muscle.
L. Vitreous humor. Optic nerve. Inferior rectus muscle. " " O. Superior P. Levator palpebrse muscle.
H.
/.
Sclerotic coat. Choroid coat-
Meibomian glands.
teaches, also, that
an image
of a certain size
produced
implies
by an
definite
object
at
a given distance
always
magnitude; and we are therefore able to judge

size of
something about the actual

size of its
an object from the

its
image and our knowledge of
remoteness.

2.
Function of the Microscope.
In order that a visual

ordinarily be invisible.
image
retina
may be
produced, a certain definite area of the
must be stimulated, and objects whose images are
smaller than this
minimum wiU
Normal
vision
is
therefore limited to fairly large objects

If,
near at hand.
however, the rays proceeding from
more remote, or
smaller, objects can be collected
by a
lens and bent so as to produce a larger image, the objects
may be
of large
seen.
This end
is
attained by the telescope
and
the microscope,
the
former producing enlarged images
and remote
of
objects, the latter producing enlarged
images of near and minute objects.

3.
Laws
Refraction.
^The
is
formation
of
such
images by the microscope

that rays of light
of
dependent upon the fact
in passing from any
medium
to
one
different density
experience
a change in direction,
unless
they
impinge at right angles to the surface of
contact.
The
deflection thus
produced has been compared
to
the alteration in the course of a column of troops on pass-
ing from a smooth parade-ground into a ploughed

If,
field.
in Fig. 2, ahafii represents a
body
of troops
it is
march-
ing in the direction indicated by the arrow,

that
apparent
the
men on
field
the right of the line will reach the

first
ploughed
tarded.
bâ^bâ,
and
will
be somewhat
re-
The
left of
the line will gain
upon the
right
and
the
column
Hne
as
as a whole will execute a partial right face.

field
In passing out of the

of the
will
it
on the other
side,
first
the right
reach the smooth ground
and gain
as
much
lost before, the final result being that the

in a direction parallel to that
column proceeds
which
it
originally pursued.
It will
be noted that in passing into the region offering
greater resistance the marching

tion
column takes up a
direc-
more nearly
at right angles to the

it is
boundary of that
line
region; in other words,
deflected toward a
drawn
So
normally, or at right angles, to that boundary
(}l^.
Fig. 2.
Îllustration of Refeaction.
(After
Hager-Mez.)
when a ray of dense medium
light passes
it
from a
less
dense to a more
is
always refracted toward the normal,
while in passing from a more dense to a less dense

it is
medium
bent away from the normal.

Refraction in the
4.
Convex Lens.
ray of light
passing through a piece of glass with parallel sides will
simply be shifted
laterally, as in
our illustration of the
marching column.
ever,
In passing through a prism, how3.
a resultant bending ensues as shown in Fig.
FUNCTION AND PARTS OP THE MICROSCOPE.
Ray AB, bent toward

and, bent
the normal at B, turns to the right

the normal to the surface, at C,
further.
If
away from
still
turns to the right
two such prisms are
Fig.
3.Refraction by Prisms.
placed base to base as in the figure, rays striking them, as
do the
lines
AB
and AB',
will
be bent in opposite direc-
tions so that they will
meet
at the point
D.
The amount
of bending varies with the angle at the apex of che prism.
As shown,
for example, in Fig. 4, the ray a'h'c'd' passing
Fio. 4.
^Refraction
in Prisms or
Differfnt Angles.
through a prism of wide angle will suffer greater

tion than the ray abed
is
deflecit
which meets surfaces
to
which
more nearly normal.
A
up
biconvex
lens
works roughly, as
of prisms
if
it
were made
of
an
infinite
number
arranged about a
central axis.
Rays
parallel to the principal axis of such
a lens (the
line joining the
centers of curvature of
all
its
two
that
surfaces,
axis,
will
be bent from
directions
toward
or
and the rays nearest the outer edge

all will
periphery will be most bent, so that

point,
meet
at
one
known
(o.
as the Principal
Focus or burning-point of
the lens
Fig. 5).
The
greater the curvature of the
Fig. s-
^Plano-convex Lens and its Principal Focus.

(After Hager-Mez.)
its
lens the nearer to

will
5.
lie.
optical center this principal focus
Formation of Images by the Convex Lens.

is
Âs
the principal focus of a biconvex lens
defined as the
point at which rays parallel to the principal axis meet,

it
is
obvious that rays proceeding from a radiant point
placed at the principal focus will be sent off on the other

side of the lens as parallel rays,
and and
will
meet
Fig.
to
form an
image only
at
an
infinite
distance
lens
{B,
its
6).
Rays
from a point between the

being
still
principal focus,
more
divergent, will not even be
made
parallel,
but will
still
continue to diverge after passing through.

the lens, though of course less so than before {A
,
Fig. 6)
No
real
image can therefore be formed by an object
lying inside the principal focus.

side the principal focus
Rays from a point

initial
out-
have an
divergence so small
as to be entirely overcome by the lens.
Such rays (C,
Fig.
6.
Course of Rays according to the Relation of a Luminous PonsTT to the Principal Focus.
vîll
Fig. 6) lens
actually converge
afteir
passage through the
and
will
meet
at
a definite point to form a real
image.
6.
Construction
of
Real and Virtual linages.The

is
construction of the image formed by any object

simple,
if
very
only
its
relation to the principal focus of the
lens be determined.
The
course of two rays from any
point
is
known
with certainty.
The
ray parallel to the
principal axis will be so bent as to pass through the principal focus
on the opposite
side of the lens.
The
ray
passing through the optical center of the lens will not be

all,
deflected at
because
it
will cut surfaces
which are
parallel to each other.
Where
lens
these two rays meet the
image
is
of the point
must be formed.
Thus
in Fig.
shown a biconvex
the surfaces of which
have
lie
an equal curvature;
equal distances from
its
principal foci, therefore,

center.
at
its
The
object ah
lies outis
side the principal focus F.
The image
of the point a
Fig.
7.-
-Formation of Image by Object outside the Principal

Focus.
(After Hager-Mez.)
determined by the straight line aa^ and the broken

aca^.
line
The image
will
of the point h is similarly fixed,
and
a'h'.
between the two

This
will
be formed the enlarged image
be a real image
one, that
is,
which could be
caught upon a screen held in the right plane,
and
it
will
be inverted.
These are the
characteristics of all images

focus.
lies
formed by objects lying outside the principal

Fig. 8 illustrates the other case, in
inside the principal focus.
which the object

it
Here
is
evident that the
rays from the point a will not meet at

side
of
.
all
on the opposite
the
lens
and
therefore
no
real
image can be

formed.
point
9
They
will
appear to the eye

their
to
come from a
and the
This
prolonging
more remote than

and
true
position;
exact location of this point

the lines from o
may be found by
backward
until they meet.
Fi(5. 8.
Formation of Image by Object inside the Principal

Focus.
(After Hager-Mez.)
image
will not
be inverted, but
it is
erect;
and
as
it
cannot be
caught upon a screen

7.
called a virtual image.
Development
of the Simple
Microscope.The
effect
of globes of crystal in concentrating the sun's rays

single point
on a
was known
in very early times.
Aristophanes,
Pliny the Elder, and other Greek and
Roman
authors
Sen-
mention
the use of such primitive burning-glasses.
eca states that "letters though small and indistinct are

seen enlarged and more distinct through a globe of glass
filled
with water."
None
of the ancients appear,
howthis
ever, to
have thought of any practical application of

as an aid to- vision;
phenomenon
up
and medical
writers
to the thirteenth century of the Christian era
speak of
short-sightedness as an incurable infirmity.
In microscopy, as in so
many
other branches of knowl-
lo

first
edge, the
distinct
advances were made in Arabia.
Alhazen, the great Arabian physician of the eleventh

century, distinctly describes the use of lenses for pro-
ducing enlarged images, in a publication supposed to

date
from
about
1052.
Two
effect.
centuries
after,
Roger
Bacon, the
Franciscan monk, and noted
alchemist, of
Oxford, noted the same
little
later,
near the
applied
Salvino
end
of the thirteenth century, lenses were
first
to the mitigation of defects of vision;
and
to
d'Armato
spectacles
degli Armati,
is
a Florentine, the invention of

use of lenses as micrdscopes
ascribed.
The
for the examination of objects too minute to be studied
with the unaided eye became general about the end of

the sixteenth century.
From
the year 1600 such obserfirst
vations were numerous, and in 1637 the
diagram of
a microscope no~w extant was published by Descartes.
About 1665 small
glass globules
began
to
be used instead
of convex lenses for the simple microscope.

set in
They were
metal plates, on the side of which opposite to the
observer the object to be examined was mounted on some

sort
of
movable arm.
With
these instruments a high
magnification was
obtainable;
and
it
was with such
simple microscopes (Fig. 9) that the pioneer microscopists

of the seventeenth century, Kircher in Italy
and Leeuwen-
hoek
8.
in Holland,
founded the science of Micro- Biology.

Microscope.
The Compound
was
Meanwhile
step
had been taken, which, though not

at the time,
to
particularly fruitful
become
the
later of great significance.
This was the invention of the compound microscope
commonly
attributed
to
Dutch spectacle-makers.

FUNCTION /iND PARTS OF THE MICROSCOPE.
Hans and Zacharias

about 1590.
Janssen, and supposed to date from
It is certain that
compound microscope
in 16 10,
was independently invented by Galileo

Cornelius
and that
Drebbel in Holland was credited with the
introduction of the instrument about 1621.
These
early
Fig. 9.
Leetjwenhoek's Microscope (Circa

(After Carpenter-Dallinger.)
1700).
instruments and their illuminating apparatus were cum-
brous and unwieldy in the extreme (see Fig. 10).
The compound microscope

that
it
is
characterized by the fact
contains two or more lenses or systems of lenses,
one of which forms an image of the object, while the other

forms a second image of the
first
is
image.
The
course of
the rays in such an instrument

object
shown
in Fig. 11.
The
^5 lies outside the principal focus of the lens system

Objective,
marked
at
and
its
is
real inverted
at
image
is
formed
A ^B^.
This image
produced
a point inside the
principal focus of the Ocular or Eyepiece;
and the
eye-

12

and
virtual image,
piece forms an erect
A^B^, of the real
inverted image
g.
^B^.
Defects of the
Compound
Microscopes.
^The
com-
FiG. lo.
Hooke's Compound Microscope

(1665).
pound microscope was

tance,
at first of
little
practical impor-
on account
of
the complications introduced by

aberration.
spherical
and chromatic
The former
is
due
to the fact that the
rays which pass through the outer
13
Fig. II.
CotTRSE of
Rays in the Compound Microscope

(After Gage.)
14
ELEMENTS OF
/APPLIED MICROSCOPY.
part or periphery of a biconcave lens are brought to a focus
somewhat
closer to the lens than those
which
lie
nearer
the optic axis.

of the
This deflection and the consequent blurring
image increase with every increase in the curva-
ture of the lens.
still
more
serious defect lies in the
fact that the rays of light of different colors are differ-
ently affected
by the ordinary
lens, those of shorter

,
waveto a
length at the violet end of the spectrum, (v)
coming
focus
first,
as indicated in Fig. 12.
The
result is that

(in virtue of
15
which white
light is split
up
into
its
compo-
nents) varied independently of each other.
Thus flint glass

glass
If
has only
.1
to .2
more
refractive
power than crown
with more than twice

lens of
its
dispersive power.
,
a biconvex
crown
glass, (C)
be combined with a plano-con,
cave lens of
as
flint glass, (i^)

it is
arranged in the opposite sense,
shown
in Fig. 13,
possible so to adjust their oppoflint glass
site
all
curvatures that the
shall
compensate for
the dispersion caused by the crown glass
and neu-
Fio. 13.
^The Achromatic Objective.
(After Hager-Mez.)
tralize
only half
its
refraction.
This
is
the principle of
efforts
the
achromatic objective;
and through the
of
Selligues
and Chevalier
in Italy,
in France,
Fraunhofer in Gerin
many, Amici
England,
it
and Goring, Tulley, and Lister

its
gradually attained
practical development
this
between 1820 and 183b.
With
improvement the
compound microscope acquired new importance;

it
and
very shortly developed into one of the most important
instruments at the disposal of modern science.

II.
The Immersion
flint
Objective.
Lenses made of combe further reduced to a
bined crown and
glass in such fashion as to be
achromatic will also show a decreased spherical aberration,
and
this latter defect
may
minimum
by adjusting the radii of curvature of the oppo-

i6
ELEMENTS Oh APPLIED MICROSCOPY.

surfaces of the lenses according to certain
site
known
relations.
Such lenses are designated as aplanatic.

still
There
remained one serious limitation
to the in-
crease of the
power
In
of magnification of the
compound
is
microscope.
of light,
ture,
all its
various refractions there
loss
and with the small objective
lenses of great curvait is
necessary for high magnification,
difficult to get
a sufficient illumination for clear vision.

it
Furthermore,
has been shown by Abbe and others that the rays

of
which extend from a point toward the periphery

lens
a
of
are
finer
of
prime
importance
of
objects.
of
in
If
the the
detection
the
structure
lost,
outer zones of
details
rays
are
no image
this
is
very minute
can
be formed; and
just
what occurs
is
in
air
the ordi-
nary compound microscope when there

the objective
between
is
and the specimen
to
be examined, as
If,
shown
in the right half of Fig.
14.
on the other
Fig. 14.
Epfect or Homogeneous Immersion.
(After Hager-Mez.)
hand, some substance
like cedar-oil,
which has the same
refractive index as glass,
be placed between the lens and
the cover-slip which covers the specimen, the rays will
17
take the direction indicated on the left-hand side, more

peripheral rays will enter the lens,
will
and more
detailed images
first/
be produced.
Such an immersion objective was
suggested by Amici in 1850, and next to the achromatic

objective this
in the
may
be considered the greatest single step

of the
improvement
compound microscope.
The
of light
degree to which a lens admits the peripheral rays

is
designated by the term angular aperture, which
signifies the angle
contained between the most divergent
rays passing through the objective from the axial part

of
an object
(a point situated
on the principal
axis of
the lens).
Obviously the angle will increase with the

its
convexity of the lens and

tance
;
consequent short focal
dis-
with the same lens, the angle will be greater
when
some homogeneous immersion substance is used. Taking this factor into account, the power of an objective to
collect
ical
and
utilize divergent light-rays is called its
Numerby
Aperture;
this quantity is
equal to the index of re-
fraction of the
medium
in front of the lens multiplied
the sine of half the angle of aperture.

12.
The Mechanical Parts
of
the Microscope.
The
and
microscope
lenses of
consists in its essentials of the
two systems of
which we have spoken.

needed
For steadiness these
optical
parts must be mounted

is
ilpon a rigid stand;

for
in addition apparatus
throwing light upon
the object to be
examined and
for focusing, or so ad-
justing the relation between the object and the

that a clear image
lenses
may
be produced.
illus-
microscope of the ordinary American pattern,

in
trated
Fig.
15,
has a heavy hors&shoe-shaped
base

i8
ELEMENTS OF APPLIED MICROSCOPY,
Fig. 15.
^The Microscope,
j^'
L,.
f
A. Base.
B.
C.
Pillar.
Coarse adjustment.
Arm.
Fine adjustment.
Stage.
clips.
D. Tube. E. Collar. F. Objectives.

G. Ocular.
M.
O. P. Q. 5.
N. Spring
Mirror.
H. Draw-tube scale. 1. Draw-tube.
Mirror bar. Substage.
Diaphragm.

and a
vertical .pillar rising
19
from
it,
which may or may

the stage, a
not be jointed.
perforated plate
Attached to the
pillar is
upon which
the object
may be
placed;
is
below
this is the
Hghting mechanism, and above
the
arm
carrying the optical parts.

includes
first
The Hghting apparatus

on a jointed bar as
it
a mirror, usually
having one plane and one concave surface and so mounted

to set at
any angle.
For high powers
is
customary to concentrate the Ught from the mirror

of
by means
below the
an Abbe condenser, a large lens placed
stage.
is
The
stage itself in the simpler microscopes
merely a
fiat plate
with an opening in the center, and with clamps
for holding the shde
upon which the object

is
is
mounted.
For deHcate work
it
convenient to use a microscope
equipped with a mechanical stage which can be moved

forward and backward and from side to side by a micrometer
screw.
Such an arrangement makes
it
easier
to
explore the whole of the specimen to be examined, and
enables one to find any particular portion of

ure.
it
at pleas-
Under
for
the stage, whether
it
be of the simple or
the mechanical type, there should be some sort of dia-
phragm
will
regulating the
direction
from which
light
reach the object, and cutting
off the peripheral rays
coming from outside the object itself which tend to obscure the image. The diaphragm opening must therefore vary with the size of the object under examination
and with the power
of the microscope, a smaller opening
being used with a higher power.
This adjustment
is
accompHshed by a revolving
disc with openings of dif-

size
ferent
in
the
cruder microscopes, and by an Iris
diaphragm
in
is
more elaborate instruments.

borne by the upper part of the
pillar
The arm
moves
adjustment,
which
in the lower part,

is
and
this
motion, called the fine
regulated by a milled head at the top.

the distance between the lenses
By
sive
this
means
and the
extenpossi-
object
may be
varied with great delicacy.
More
changes in the position of the lenses are
made
ble up'
by the
fact that the tube in
which they are fixed sKdes

it
and down
in the
clamp which holds
at the
end of
the arm.
This movement, called the coarse adjustment,
may be regulated by hand or by a rack-and-pinion. The tube itself is divided into two portions, the
proper and an inner cylinder, the draw-tube, which
tube
may
Into
be pulled out telescope fashion from
its
upper end.
the lower end of the tube proper the various objectives
may
is
is
be screwed, while the eyepieces or oculars
slip into
the upper end of the draw-tube.
When
the -draw-tube
pulled out the space between the two systems of lenses

increased,
and
in order that the real

visible
image produced
it
by the objective may be
through the eyepiece,
must be formed farther up than would

case.
ordinarily be the
that, according to
A reference
to Fig. i6 will
show
the principles previously deduced, this can be accom-
plished
by bringing the object nearer the principal focus

and that the image
of the object so
of the objective,
brought nearer the principal focus will be correspondingly
enlarged.
Thus puUing out

sets
the
draw-tube
in-
creases the magnification.
At
least
two
of lenses
accompany a compound

microscope,
21
commonly two
eyepieces
and two or more
objectives of different magnifying powers.
The
lenses of
some makers are designated by

rule,
arbitrary symbols;
as a
however, they are marked with numerals w;hich
indicate
what
is
known
as their equivalent focus, or the
focal length of a simple converging lens
which would
Fig.
1 6.
Relation or Distance from Principal Focus to Size AND Position of Image.

size as that
produce an image of the same

the lens in question.
formed by
The larger the number the lower Thus a i|-in. eyepiece magnifies less than a i-in. eyepiece, and a J-in. objective less than The commonest high-power objeca ^-in. objective.
will
be the power.
tive is the
i^j,
and the f 5
is
the highest objective which
can be practically constructed.

violet rays with
By making
use of ultra-
a wave-length only half that of ordinary
light the Zeiss Optical
to construct instruments of
Company has recently been able much higher power. The

and the image
recorded on the
invisible,
is
lenses in this case are of fused quartz,
produced, being of course
photographic plate.
22
ELEMENTS OF /IPPLIED MICROSCOPY.

In some microscopes the two lenses of the eyepiece
work together
to
produce the
effect
shown
in Fig. ii.
is
More commonly, however,

formed between these two
one forms the
virtual
the
real
inverted image
lenses,
and only the upper

such
image.
In
an eyepiece,
known
as a negative or
Huyghenian
ocular, the lower
or field lens really forms part of the objective system.
REFERENCES.
Batjsch, E.
Manipulation of the Microscope.

B.,
Rochester, 1901.
and Dallinger, W. H. The Microscope and London, 1901. its Revelations. Clark, C. H. Practical Methods in Microscopy. Boston, 1894. Gage, S. H. The Microscope. Ithaca, N. Y., 1904. Hagee, H., and Mez, C. Das Mikroskop und seine Anwendung.
Caepenter, W.
Berlin, 1899.
These
the
references,
and those appended
to other chapters, include
only those few books which have been, found most indispensable in
work
of the course in Industrial
Microscopy as given at the Inbibliography of works on
stitute
of Technology.
will
very
full
microscopy
be found in Professor Gage's book cited above.
CHAPTER
II.
THE MANIPULATION OF THE MICROSCOPE.

I.
Setting
Up
the
Microscope.
The
microscope
is
an instrument
of precision
whose deUcacy may be
easily
impaired by carelessness or neglect.

it
When
it
not in use
in
its
should be protected from dust by placing

bell-glass,
case
settle
or under a
and any
particles
which
upon
it
should be removed with a camel's-hair brush

If
and chamois-skin.
necessary,
the mechanical parts
may be
first
cleaned with chamois-leather moistened in a
solution of equal parts of benzine
and
olive-oil
and then
wiped with dry chamois.
The microscope
should be handled always by the base,

of the pillar, in order to avoid
and not by the upper part
straining the fine adjustment.
When
who
in use
it
should be
placed on the work-table rather near the edge, with the

pillar side nearest the observer,
should
sit
close to
the table in a chair of such height that his eyes will be a

little
above the
Fig.
17).
sit
level of the
upper end of the draw-tube
(see
Those unfamiliar with the microscope

For general work
23
are apt to
too far off or too high above the instrument

it.
and bend painfully toward

best not to
tilt
it
is
the tube of the microscope even
when
the

24
pillar is

provided with a joint for that purpose;
is
and
if
the instrument
kept upright, the spring clips
may
be
the
dispensed with, the object being

stage as desired.
moved about on
Fig. 17.
Proper Position for Observer.
(After Bausch.)
The
lenses of the microscope should always be

if
exam-
ined before they are inserted in order to see

quire cleaning.
soft clean cloth
they re-
Since the glass

or,
is
easily scratched, only

is
better
still,
Japanese lens-paper
suitable for this purpose;
the fingers, which are always
MANIPULATION OF THE MICROSCOPE.

oily,
25
must never be allowed
to touch
the lens.
Well-
defined spots seen on looking through the microscope at
a clear
as
field are
due to specks of dust upon the eyepiece,

rotating that system of lenses.
may be proved by
Not
infrequently such specks are formed on the upper surface of the lower or field lens of the ocular,
which must
be unscrewed in order to remove them.

ness
ive;
is
Diffuse cloudi-
generally caused
this
it
by
dirt or
moisture on the object-
and
may
be removed by breathing on the glass
and wiping
smears of
with lens-paper.
Sometimes for stubborn

alcohol or xylol
dirt or grease,
95%
may be may
not
used, but in the latter case the solvent must be sparingly
applied and proonptly removed in order that

affect the setting of the lenses.
2.
it
Lighting.
Âfter the objective
has been screwed in

in Fig. 18,
at the
bottom of the tube, as shown

at the
and
the
eyepiece has been inserted

tube, the mirror should
available illumination.
upper end of the draw-
be
so adjusted as to give the best
Direct sunlight must of course

fur-
be avoided, but a clear area of sky or a white cloud

nishes an
ideal source of light;
and sunlight
directly
reflected
from a white wall or transmitted

tain
through a cur-
may be
used to advantage.
For
artificial illumina-
tion electric bulbs with ground-glass shades are suitable,
or the narrow edge of the flame from a fiat-wicked
oil-
lamp may be
satisfactory
substituted.
gas-flame,
unless
some
is
incandescent mantle be attached to the burner,
not
on account
of
its
unsteadiness.
The
flat
surface of the mirror yields sufficient light
under ordinary circumstances, but the concave surface

26
may be used when the strongest illumination is desirable. The observer should so adjust its angle in relation to the
window
light
or
lamp
seen.
that a clear
and well-defined
circle of
may be
If the light
be uneven, or
if
an image
of the
window-bars
or the flame be visible, either the angle of the mirror or

its
distance from the stage
may be
so
changed as
to re-
Method
of Inserting Objectives.
(After Bausch.)
move
the difficulty.
If the circle is too bright so that its
glare hurts the eyes, the angle of the mirror
must be
changed
to
moderate
it
and the diaphragm opening

desirable that the Ught should
should be decreased.
For most purposes

pass directly
it
is
upward through
This end
the axis of the microscope
axial illumination.
may
be attained by focus-
ing just
below an air-bubble, and so arranging the mirror
that the bright point inside shall be exactly at the center of the bubble.
In examining diatoms and some other
27
objects, oblique illumination brings out certain structures

best.
Light so oblique that

is
it
cannot enter the objective
at all
sometimes obtained by placing under the stage

off the
a stop cutting
central cone of light.
In such a
preparation the object will be
made
visible
by the rays
which
it
reflects
or refracts upward, and will appear
self-luminous
illumination.
on
black
background
dark-ground
With objectives higher than J in. even the concave mirror will not give sufficient light for successful microscopic work, and the
the plane mirror.
Ahh6 condenser must be Used, with The condenser, as has been stated in
of lenses placed just
to concentrate k
|
Chapter
I,
is
an objective system
under the stage in such a position as

considerable
amount
of light
upon
the object to be
ex!-
amined
(Fig. 19).
With low powers the condenser should

its
be swung out from
place to one side;

results
while with the
i^-in. immersion objective the best
may be obtained
by placing a drop
bottom of the
homogeneous.
magnifications
of oil
between the condenser and the

entire system optically
slide,
making the
one
is
When
it
working with these high
is
also
important that the condenser

that
is,
should be accurately centered and focussed;
it
must
be at such a distance below the stage that the great-
est possible
amount
of light
the object.
This position
may be concentrated upon may be determined by fdcus-
ing upon some specimen with a -in. objective and then

so adjusting the condenser that the image of a window-
sash or of a flame coincides with the object

It
itself.
has been pointed out that the function of the
dia--

28

is
phragm
finally
to cut off adventitious light so that all the rays
reaching the
microscope shall come from the
immediate
will
vicinity of the object.
A little
experimentation
show
that reducing the
diaphragm opening often
gives a sharper picture as well as one

to the eyes;
much
less trying
in general
an opening about the

good
size of the
front lens of the objective will yield
results.
Fig. 19.
Âbbe Condenser.
(After Hager-Mez.)
Certain opaque objects are best examined by reflected

light,
and
in
many
no
cases this
method may well be used
to
supplement the ordinary one, the mirror being turned

aside so that
light passes
through the stage and the
object being illuminated either with the light which naturally falls
on
it
or with rays concentrated from above
by a
3.
lens or mirror.
Focusing.After the
is
field is well lighted, the
next
step
to focus
on the
object, or to so adjust the relation
pf the lens systems that a clear image
may
be formed.
29
The low-power
for preliminary
objectives
(those of one-half inch
and
one-third inch equivalent focus) should always be used
exploration before a
new specimen
is
examined with the higher powers,

of view
since their large field
makes
is
the general relations of the object clear.
Focusing
also
much
easier with the lower powers, since
with them a small object

their
tive
may
easily
be found, while
working distance, or the space between the objec-
and
the object
when
in focus,
is
great enough to
allow free play.
After placing the object over the center
of the opening of the stage, the tube should
be run down
is
by means
of the coarse adjustment until the front lens
within an eighth of an inch of the object.
Then, looking
through the microscope, the tube
is
slowly raised by the
coarse adjustment until the object comes

clearly into view.
It is often helpful to
more or
less
move
the slide
about with the

outline of a
left
hand
at the
is
same
time, as the
shadowy
moving object
more
readily recognized than
that of one at rest.
As soon
as the object
is
dimly seen
it
clearly into focus
by the use
of the fine adjustment.

is
may be brought The

of great im-
proper manipulation of this mechanism

portance, since
if
the attempt be
made
to study objects
which are not well
in focus, the eyes will be strained
and
structures incorrectly seen.
Furthermore, at any given
position of the fine adjustment only a certain plane of the
object examined will be in focus, while
its
whole figure
is
needed to make up a correct
picture.
Therefore, one
fine
hand should be kept continually on the

while the microscope
is
adjustment
needs
in use, varying
it
slightly as

It
demand.
fine
should be noted that the excursion of the

is
adjustment
it
necessarily small;
its
if
care
its
is
not taken
will
to
keep
near the middle of

it
range,
motion
be
its
stopped short at one end or

place at the other.
will
be unscrewed from
Focusing with the higher power
is
more
it
difficult.
On
account of the short working distance

"run the objective
is
necessary to
down
as close to the object as possible,

side,
watching
its
approach from the
and then focusing
up very
carefully with the fine adjustment.
Useful practice in focusing
may
be obtained by the
study of
air-bubbles
and
oil-drops.
few drops of
clove-oil should
be mixed with mucilage and the mixture
beaten up on a slide with a knife-blade so as to insure
MANIPULATION OF THE MICROSCOPB.

the ring widening the
this
31
and the
center
becoming sharper as
objective
is
passes
downward.
With the oil-bubble

and the
widest,
reversed,
the brightest
center
is
ring being above.

Fig.
18.
Why
like
this
so will
be seen from
less
The
oil-bubble,
a dense sphere in a
dense medium, acts

light to
an ordinary lens concentrating
a bright focus above, the surrounding ring from

light is diverted being dark.
which the
will
This dark ring
widen upward.
less
The
air-bubble,
on the other hand,
being
light, 4.
dense than the mucilage, diverges the rays of
and its -dark ring is widest below. The Use of the Draw-tube. The
spherical
its
and
rela-
chromatic aberration of an objective vary with

tion to the object fore
examined and
to the eyepiece.
There-
when we
say that in an achromatic objective these

this is only true for
defects
have been corrected,
certa,in
standard set of conditions.
If
we change
the position of
the draw-tube and therefore the distance between the

objective
and eyepiece
systems, a certain
amount
of aberis
ration
is
again introduced.
For each objective there
certain proper tube-length, record of which accompanies
the microscope
itself.
and may be placed upon the

to
objective
According
the
best
American
practice
the
tube-length,
measured between the upper end

is
of the tube
where the eyepiece
inserted
is
and the lower end

inserted,
is
of the
tube where the objective

216
either
160 or
mm.
cover-glass
is
A
and
commonly placed between

and
this introduces
the object
and the
objective in order to hold the former in place
to protect the latter;
a pertur-
3*
ELEMENTS OF APPLIED MICROSCOPY:

for.
bation which must be allowed

21 will
Reference to Fig.
cover-glass shifts
arise
show
in a general
way how a
the rays coming from a
point,
F, and makes them
apparently from F' and F".
In the making of achrotaken into
matic
objectives this effect of the cover-glass is
account;
but with each
lens,
standard results can only
be obtained with cover-glasses of a certain thickness.

variation of .05
mm.
in thickness
may
quite obliterate
Fig. 21.
DiSTtTRBiNG
Effect of the Cover-glass.
(After Gage.)
certain fine structures.
Adjustable objectives are so
made
that
by turning a
ring or collar the. distance between
their systems of lenses
may be may
is
varied, being increased
for the thinner cover-glasses.
With an ordinary unadselect
justable objective
one
it
cover-glasses
of the
thickness for which

easily
corrected, the dimension being

is
measured by some such apparatus, as
shown
in
Fig. 22.
Or
the lens system
may be
by
adjusted for cover-
glasses other than the standard
altering the position of
the
draw-tube,
changes in the
tube-length
producing
changes similar to those which are provided for in the adjustable objective.
for thin covers
The
tube-length should be increased

for those thicker than the
and decreased

MANIPUL/ITION OP THE MICROSCOPE.
standard.
33
Such changes
in adjustment are necessary only
in the case of dehcate work.

5.
Care of the Eyes.
Much
of the
difficulty
some-
times experienced by beginners with the microscope
may
be avoided by attention to three points mentioned above
the cleanliness
of the lenses, a clear but not excessive

field,
illumination of the
and a proper manipulation
of
the fine adjustment.
In no case should the observer
strain his eyes in the attempt to study
what he cannot
Fig. 22.
Micrometer Calipers.
see clearly.
If the object is not distinct, there
must be
something wrong which should be remedied.
The
distance of the eye above the eyepiece should
vary with the magnification.

lenses there
is
For each combination of
a certain eye-point at which a

is
maximum
and above
number
shadows
of rays
most
closely concentrated,
field will
or below, the size of the

will appear.
be reduced, and colored
The
higher the power the nearer
the eye-point approaches to the eyepiece.

It is best to
accustom oneself
to the use of
both eyes
alternately,
and
to acquire the habit of keeping
open the
eye which
is
not over the instrument.
At
first
the atten-
34
tion
may
be distracted by external objects, but
this diffi-
culty will pass with practice.
The beginner may
also be
misled by seeing through the microscope certain cloudy

specks floating across the
field of view.
These are the
muscae volitantes, shreds of matter lying in the vitreous
humor
of the eye; after a time they are so discounted

their
by the observer that he becomes unconscious of

presence.
6.
Qualifications of a
Good Microscope.
The
princi-
pal parts of the
compound microscope have been

it
de-
scribed in Chapter I;
of the
remains only to point out some
most important
qualities
by which we may measure

Differences in
the value of any individual instrument.
the mechanical parts are largely a matter of personal

preference, although the stand should be as
compact as
possible with a base sufficiently heavy to give proper

steadiness.
Both the coarse and
fine
adjustments must
work
slip
easily without being so loose as to allow the tube to
down
of its
own
weight.
have
an ample excursion.
freely,
The fine adjustment should The mirror-bar ought to

any position
in
move
and yet
retain
which
it
is
placed.
The
stage should be wide
enough
to
accommo-
date large objects
when
necessary.
The
of
optical parts are, of course, of prime importance;
they should be examined with respect to the four quahties
magnifying power, resolving power, penetration, and
illuminating power.
ratio
The
magnification of a lens, or the

size of the
its
between the
size of
an object and the
image
formed, depends simply on the curvature of
surfaces
in
and may be measured by the method described
Chap-

MANIPUL/ITION OF THE MICROSCOPE.
ter
35
IV.
The
resolving
power by which
fine structures are
made
as has
visible varies directly with the
numerical aperture,
is
been already explained.
This power
ordinarily
tested bytera or
examining the wing-scales of certain Lepidoppossess very fine
the shells of diatoms which
markings.
For example, the
shell of
Pleurosigma shows
three systems of striations

nification of
when examined under a maglesser aper-
250
diameters with a numerical aperture
of over .80, while

ture.
two of them disappear with
Penetration, or the power to see clearly different
planes of the object at the same time, varies inversely
with the numerical aperture and directly with the square

of the equivalent focus of the lens.
Illuminating power
varies with
the square of
the numerical aperture,
and
with the square of the equivalent focus.
Two common
faults should be looked for in a micro-
scope, curvature of the field
and imperfect correction

If a stage
of
chromatic and spherical aberration.

ter,
microme-
marked with
lines at right angles,

a, Fig. 23.
be examined, the
it
image should resemble
If
has the appear-
Fig. 23.
Curvature of the Field.
(After Hager-Mez.)
ance of
tion
is
&
or c the lenses are so ground that the magnifica-
greater or less at the periphery than at the center.
36
Spherical aberration
of the test objects
may
be detected by examining one.

If the lens
mentioned above.
be under-
corrected, so that peripheral rays
come
to a focus nearer
the lens, the outer part of the object will be in focus in a
plane below the central portion.
Chromatic aberration
may
7.
be detected in the examination of any object by the

it.
appearance of colored rings surrounding

Interpretation of Appearances.
in
The
greatest care
to
must be taken
structure
of
drawing conclusions as
the real
bodies from their appearance
under the
invert-
microscope.
Right and
left
are reversed
by the
ing action of the objective lenses.
Appearances in any
air-
one plane
bubble at
object
is
may
its
be very deceptive, as in the case of an
upper edge.
The medium
III.
It
in
which an
mounted may completely

Chapter
alter its appearance,
as
we
shall see in
must always be
re-
membered
that contrasts of density are
what give us mi-
croscopic pictures;
only
when
these coincide with salient
differences in structure will such pictures be representative.
The
presence of dust or other foreign materials
may
mislead the observer, and even air-bubbles have

If the preparation
sometimes caused confusion.

treated with fixing agents,
has been
dyes,
etc.,
as described in
to mistake
Chapter
artificial
III, great care
must be exercised not
conditions due to reagents for those normally
existing.
Motion
deceptive.
of bodies
under the microscope

is
is
particularly
Since the distance travelled
magnified, an
idea of rapid
translation
movement
is
conveyed when the actual

Diffusion currents
may be
really very slow.

are often set
37
up
in the
mounting medium which may be
misinterpreted.
in the
still
more
serious source of error lies
phenomenon known
as the
Brownian movement,
or pedesis, a dancing, oscillating motion which affects
suspensions of finely divided solid particles under certain not very clearly understood conditions.
Gamboge
Brownian
or carmine
suspended in water shows the
movement
well,
and should be studied
carefully in order
that such motion
may
not be confused with that characor other living organisms.
teristic of the bacteria
REFERENCES.
Bausch, E. Manipulation of the Microscope. Rochester, Gage, S. H. The Microscope. Ithaca, N. Y., 1904.
igoi.
CHAPTER
III.
MOUNTING AND PREPARATION OF OBJECTS FOR THE MICROSCOPE.

I.
The
Effect
of
Mounting Media.
^The
Under
clearness
with which
we
see
an object depends
its its
in great part
upon
the contrast between
density
or, colo.r
and the backthe mi-
ground furnished by
surroundings.
croscope Httle remains but difference in density to emphasize outlines,
and the
distinctness of a given object will
thus vary widely according to the

lies.
medium
in
which
it
The
student
may
obtain an idea of the importance

it
of this factor
and
of the necessity for taking
into ac-
count in interpreting microscopic appearances, by the
examination of potato-starch grains in

cerin,
air,
water, gly-
and
clove-oil.
little
In
air the grains will
show heavy
black edges and
internal structure.
In water the
edges are less pronounced and the hilum and oystershell
markings appear.
In glycerin the same characin clove-oil the
teristics are still
more pronounced, and
edges are almost invisible, so that the grains have a spectral
appearance and the hila are very strongly marked.

it
Obviously,
is
necessary in making deductions as. to
the real structure of an object to consider the influence

38
MOUNTING AND PREPARATION OF

of the
OBJECTS.
39
mounting medium;
and
it
is
also desirable
by
choosing a
medium
is
of proper density to minimize as far
as possible the errors due to excessive refraction.
Thus,
when
tive
starch
is
mounted
in air, the difference in refrac-
index
so great that the heavy black edges produced

is
are most deceptive, while in clove-oil the difference

slight for clear definition.
2.
too
Temporary Mounting Media.

it
When
an object
air, it
is
of such nature that
can be well examined in
may
by
be placed upon an ordinary glass

I in., best
slide (usually 3 in.
with ground-glass edges) and examined directly
under the microscope.
Even
in this case,
however,
if
the high objectives are used, the specimen should be
covered with a cover-slip in order to protect the front lens.
Of
the cover-glasses
commonly
sold the square ones are
most convenient for ordinary handling, and the round

ones, best suited for making permanent mounts, as will
be seen
later.
The
effect of the thickness of the cover-
sHp upon microscopic vision has already been considered in Chapter II.
In
fine
work
it is
often desirable to
measure the thickness
of the covers used in order to select
those, of a standard size or to adjust the objective or
draw-tube to those which deviate from
it.
This
may
upon
be effected conveniently by means of micrometer calipers

or with certain special forms of apparatus placed
the market for
this
purpose (Fig.
22).
Most
objects are
air,
more
clearly seen in
some denser
any
liquid,
medium than
choice of a
and,
when mounted
in
cover-slip is always necessary even for low powers.
The
medium
must be determined by two consider-
4
ations.
ELEMENTS OF
First,
/tPPLIED MICROSCOPY.
its
as suggested above,
refractive
index
should be far enough, and yet not too

that of the object to be examined.
of such a nature that
it
far,
removed from
it
Second,
should be
will not set
up
destructive, os-
motic, chemical, or other changes in the substance of the
specimen.
For aquatic plants and animals and a great

fulfils
many
other objects water
these requirements;
and
for specimens of animal tissue the
normal
fluids of the
body furnish
jects,
ideal
it
mounting media.
With denser ob-
where
is
necessary to minimize contrast effects,
glycerin or
some
oil is preferable.
3. Principles
of
Permanent Mounting.
În
making
necessary
permanent mounts of microscopic objects

to
it is
guard against changes due to

of the object,
{a)
physical displace(6)
ment
ical
mounting medium, or cover;
chemor
changes in the substance or the mounting
medium
loss of the latter
by evaporation; and
its
(c)
bacterial
decom-
position of the specimen or
substratum.
First, then,
the cover-glass
must be firmly attached
to the slide either
by an adhesive mounting medium or by the application

of a special cement.
Second, the
tected that
it
from evaporation
will not
medium must be proand made of such a nature

Third, the
antiseptic.
fulfilled in three quite differ-
undergo chemical change.
mounting medium must be
These requirements are

ent ways: by mounting
"dry" or
in air, in
which case
the absence of moisture serves as an antiseptic,
by mount-
ing in aqueous media mixed with glycerin, and by mounting in balsam, a resin which sets and forms a solid mass
impervious to
all
external agencies.

4.
OBJECTS.
41
Mounting Dry or
is
in
Air.
The
method
of
dry
mounting
well suited to such objects as crystals, which
show well
in air,
and which may be dried without

It
is
suf-
fering alteration.
fairly
simple, although in this

it
respect, as well as in
permanency,
is
excelled
by the
cleaned
balsam mount. For

slides
all
methods of mounting,
first
thoroughly
Slides
and covers form a
essential.
which
have been already used and are soiled with balsam or

other resinous substances must
or turpentine.
first
be treated with xylol
Otherwise
all
slides
and covers should

of water!
this they/
be put into a cleaning mixture made up by dissolving

20 grams of potassium bichromate in 100
cc.
and adding 100
cc. of
sulphuric acid.
alcohol,
From
may be
transferred to
50%
and
in that solution!
kept until needed for use.
Both
slides
and covers should
always be handled by
their edges, since the oils of the
skin will infallibly soil their surfaces.
Given the
structed
slide
and the
cover,
there
must be conshall separate
some
sort of wall or cell
which
the two and prevent the specimen from being crushed.
The
cell
will
vary in depth with the thickness of the

cell will serve, it is
object, and,
when a shallow
it.
only neces-
sary to
directly
make a
upon
ring of cement
and place a round cover
neat
is
mount may be made by using

in Fig. 24, the sUde being
a turntable such as
shown
damped on
the
movable portion and whirled about,

shellac,
is
while a brush dipped in
marine glue.
Bell's
cement, or some similar material

the guide rings
held just over one of

tip lightly touch-
on the turntable with
42
ing the glass.
narrow, even ring
may
thus be. pro-
duced, and, by a gradual process of upbuilding, one of

considerable depth.
to
If
an object of greater thickness
is
be examined, a ring of brass or hard rubber
may be
attached to the slide by a ring of cement similarly applied.
When
the cell has
become
so dry that there
is
no dan-
ger of the cement running, the object to be
mounted may
be placed within
it
or attached to the under surface of a
Fig. 24.
^TuKNTABLE.
is
(After Gage.)
cover-sKp.
The
cover
then pressed gently

is
down on
all
the ring of cement so that contact
complete
the
way
round, and, after placing the slide on the tmrntable

is
once more, a fresh ring of cement
applied partly on
slide.
is
the edge of the cover and partly on .the
When
thus prepared the dry
mount
completed and
slide, the
it
should be labelled on the left-hand end of the
nature of the specirnen, the treatment to which
has
been subjected, the

indicated.
S.
mounting medium, and the date being
Mounting
in
Glycerin Media.
Glycerin
mounts,
bal-
although not easy to
make and
less
permanent than

sam mounts,
removing
all
OBJECTS.
43
are
suitable
it
for
microscopic plants and
other objects which
is
desirable to examine without
the water which they contain.

is
Some
placed
pre-
liminary treatment
cells
necessary even in this case, since

if
containing a large amount of water,
di-
rectly in a strong solution of glycerin,
would be torn and
distorted
,
by the
violent diffusion currents set up.

to prepare such objects
is
The
to im-
most convenient way

merse them in
(âys
io%
glycerin
and
set
them by
for a
few
under cover, as a protection from
dust, so that the
water
may
evaporate and produce a gradual concentra-
tion of the glycerin.
Specimens thus prepared may be mounted in a deep

cell filled
with pure glycerin, but

cell so that
it is
difficult
matter to
cover the
air-bubbles shall not be included,
and a
still
more
difficult
matter to cement the cover
firmly to the wetted surface of the ring.
For most pursince
poses glycerin jelly *

this
is
much more
convenient,
substance becomes solid on cooling and obviates
the necessity for a cell or for cement to attach the cover

slip.
All that
it
is
necessary in using this
medium
is
to
melt
by gentle warmth, cover the object with a goodit,
sized drop of
cool place.
6.
put on the cover-slip, and
set aside in
Mounting
in
Balsam Media.
The most permanent

is
and
*
satisfactory of all
mounting media
is
Canada balsam,
One
part of gelatin by weight
soaked in 6 parts of water

finally
7 parts of
pure glycerin are added and
i%
of phenol.
The
mixture
till it is
is
warmed
and
is
for ten to fifteen minutes, with constant stirring,
clear
then
filtered.
44
ELEMENTS OF
/IPPLIED MICROSCOPY.
since this substance sets like a rock

of refraction,
It is
and has a high index

distinct.
fir
which makes opaque objects very
commonly used
dissolved in xylol, the natural
balsam being mixed with an equal volume of the

the solution after filtering
is
oil;
concentrated to a syrupy
It
is
consistency
jelly,
by evaporation.
used
like
glycerin
being placed on the object, covered and set aside,

the
xylol
when
evaporates
and the balsam becomes
firmly set.
With an object
wing or
of firm texture, not containing water,
the preparation of a balsam

leg
mount
is
very simple.
The
be
of
an
insect, for
example,
may
easily
mounted
those
to
in this way.
With
softer specimens, especially

it is
made up
largely of water,
necessary to resort
shall
some preliminary treatment which

tissues.
harden and
dehydrate the
7.
Fixing and Dehydration.
The
first
step in the prep-
aration of a soft plant or animal tissue for balsam mount-
ing
is
to treat
it
with some agent which shall
fix
the cell
structures in exactly the condition in

in
life,
which they occurred
preventing the disintegrating changes which nor-
mally follow the death of protoplasm, and which shall

at the
same time so
act chemically
upon
the cell constituof
ents as to harden
and
protect
them from the action

later.
the
chemical agents to be applied

is
Flemming's
it
mixture
tains
one of the best of these fixing agents;
con-
.25% chromic acid, .1% osmic acid, .1% glacial

and should be allowed
to
acetic acid dissolved in water,
act for half
an hour or more.
is
saturated solution of
corrosive
sublimate
often
useful.
Absolute alcohol
OBJECTS.
45
may
be applied to the fixation of many
tissues, its action
being so rapid as to forestall the bad
effect of the violent
dehydration which
it
sets
up.
At the other extreme,

harmful
dilute alcohol (i part
90%
alcohol in 2 of water) pro-
duces
moderate
fixation
without
osmotic
changes.
After fixing with
weak
alcohol, or after
washing out
Flemming's
next step
is
fluid or corrosive
to
subhmate with water, the
dehydrate, which can best be done by

alcohol
of
treatment
with
increasing
strength.
After
the specimen has been thoroughly permeated with

alcohol,
to
is
it
30%
should be transferred to a
50%
solution, then
70%, 90%, and 95%, successively. Thus the water removed so gradually that the diffusion currents set
are not sufficiently violent to distort the tissue.
up
The
or
period of immersion in each grade of alcohol will vary
with the thickness of the specimen.
For
sections
minute objects three
to five
minutes will
suffice.
is
The
to treat
last step before mounting an object in balsam

it
with a clearing agent, that
is,
with some liquid
of high refractive index which
will penetrate its tissues
and make them

grains
clear, just
as
glycerin
makes
starchin
air.
more transparent
is
than when mounted

is
Since the object

in balsam,
it is
already in alcohol and
to
be mounted
obvious that the clearing agent should be

Cedar-oil, clove-oil,
miscible with both substances.

xylol are perhaps the
and
commonest
clearing agents.
it is
After
being treated with any one of them until

transparent, the object
thoroughly
may
be placed in balsam and
mounted.
46
The
clearing agents just described
depend
for their
action solely
upon
the fact that their high index of re-
fraction prevents loss of the light passing through the

object.
ical
In the preparation of certain very opaque botanare
specimens, clearing solutions
used which act
chemically upon the tissues and actually dissolve- certain constituents

these,
which
interfere with transparency.
Of
strong
aqueous solutions of potassium hydrate
or chloral hydrate are
commonly
used,
and they must
be allowed to act upon the specimen for several hours or

even days.
8.
Section-cutting.
entire,
Since
only very small objects can
be examined
it is
necessary in
many
cases to pre-
pare thin sections of objects for examination with the

microscope.
Even with
fibres
and
similar objects
which
can be easily studied in one dimension, cross-sections are

often desirable in order to gain an idea of their whole
structure.
Sections of rigid objects
may
be cut directly with a
sharp razor;
but few specimens are sufficiently hard to

this
be treated in
way.
In general, plant and animal
tissues are so soft that they
would give way even before
a sharp knife, and must be supported by imbedding
them
to
in
some material
of firmer texture.
it
If the sections
be cut are not very thin,
is
only necessary to surlike pith;
round the specimen as a whole by a substance

if
more
delicate
work
is
to
be done, the tissue must be

like
permeated by some material
paraffin or celloidin
cell
which
will support
each individual
wall and
make
it rigid.
OBJECTS.
47
Plant stems and leaves, of which sections a tenth of a

tniUimeter or
more
in thickness will serve,
may be
easily
imbedded and
cut in pith.
is
The
fresh tissue, without

slit
preliminary treatment,
placed in a
cut at the end of
a section of moistened elder pith.

the pith, a
the pith
left
is
A thread is tied around

Then
end
little
below the end,
to hold all together.
taken between the thumb .and forefinger of the

level with the
hand, with the forefinger almost
and the thumb lower down.

the right hand,
The
knife or razor, held in

forefinger of the left
and steadied on the

the
hand,
is
drawn toward
the pith with
body so
as to cut off a thin
slice of
its
imbedded specimen.
The
sec-
tion thus cut
may
then be examined at once in water or
prepared in dilute glycerin for a glycerin mount, or

passed through the grades of alcohol for the balsam mount.
When
bedded
this
is
thin sections,
down
to thousandths of a milli-
meter in thickness, are desired, the object must be imin paraffin or celloidin
and cut with a microtome and at the same time

paraffin,
almost
always
necessary with animal tissues,
which are
tiiore
softer than plant tissues,
opaque.
first
For imbedding
in
the object
must
be
fixed,
dehydrated, and cleared, as described
above.
It is
then placed in paraffin, kept at a temperaits
ture just above
melting-point in a suitably regulated

to
bath,
and allowed
remain until thoroughly saturated.

to
The
liquid paraffin
sort of
and the object are then transferred

cell
some
temporary
it
made
of paper, or of metal
blocks, in which
can be cooled rapidly by a current of

crystals of paraf-
nmning
water.
Slow cooling produces
'fin fatal to
clean sections.

48

Once
well imbedded, a cube of solid parafiin containis
ing the object

as
is
cut out
and
fixed in a microtome, such
illustrated in Fig. 25.
Several good instruments are
on the market,
Blake types
of
which the
Thoma and
the Minot-
may
be mentioned.
In any case the knife
and
object are firmly held in supports
moving
at right
is
angles to each other;
and one or other
of the supports
Fig. 25.
Thoma Microtome.
SO arranged that
shifted by.
by some mechanical device
it
can be
any desired amount between each excursion,

If the
according to the thickness of the sections desired.
surrounding temperature be right, the successive sections

of paraffin will adhere
by
their edges,
forming a long
ribbon.
In such a ribbon each
serial section represents
one plane in the original specimen; and the whole object
may
thus be unrolled as
it
were upon the
slide,
each
structure being easily traced
from section
to section.
Minute
selves.
It
sections obviously cannot be handled

is
by themto the
desirable, therefore, to affix
them

slide
OBJECTS.
49
this is
upon which they
are to be mounted;
and
best
accomphshed by the use
of Mayer's albumin fixais
tive.*
A minute drop of this

The
section
is
solution
placed on a clean
only the thinnest
sUde and rubbed with the finger

film remains.
till
then placed upon the shde
and gently heated
till
the paraffin just melts.
The
sec-
tion will adhere firmly to the slide
and the
paraflSn
may
unfor
be dissolved in xylol or any other clearing agent.
For objects over lo mm.

suitable, since
in diameter paraffin
is
large blocks split under the knife;
such
ding
ether,
specimens celloidin
may
be
is
used as an imbedused
dissolved
in
medium.
This
substance
and
after the evaporation of the latter is
hardened
process
it
by treatment with alcohol or chloroform.

is less
The
simple than the parafiin procedure, and
is
not
possible to cut such thin sections of the objects imbedded.
The
freezing microtome, in which the water in the

its
specimen acts at a low temperature, as

substance,
is
imbedding
Cell
rapid and yields very thin sections.
structures are, however,

ess,
somewhat
distorted in this proc-
and
its
use
is
confined mainly to the preparation of
pathological material.
9. Staining.
One more
process in the preparation of

still
objects
for
the
microscope
demands reference
Since the elements
it is
the process of differential staining.
of a tissue or of a cell differ in chemical composition,
possible
to apply
certain
dyes which shall enter into
combination with some of them and not with others,

* 50
erin
cc. of
I
the albumin of hen's eggs
is
and
gram
of sodium salicylate, shaken well
mixed with 50 cc. of glycand filtered.
5-

their
and thus emphasize

ture
outhne and bring out the strucstain
more
clearly.
Sometimes the
it
acts
upon a
certain tissue as a whole, picking
out from other tissues;
surrounding
it.
More
generally, however, the substances;

cell,
used act on a certain part of the individual

the nuclteus,
generally,
clearly;
making
that
structure
stand out
from the cytoplasm.

Staining processes are often complicated, and include,
not only direct methods in which the stain

act just long
is
allowed to
enough
to affect the desired elements and;
then washed out, but also indirect methods in which the

tissue is overstained
and the dye then removed by

it
alco-
hol or acid from the parts which give
up most
,
readily.-
Small objects
treated
may
slide.
be. stained
in
bulk and section^
on the
Of
anilin
the
staining
solutions
used,
hamatein and the

eosin,
dyes
(fuchsin,
etc.)
methylene blue,
are the
safranin,
useful.
Bismarck brown,
most generally
it is
By
a proper combination of two of these
possible to
stain the cytoplasm of the cell
one color and the nuclei
another.
This process of double staining depends of
course on the greater avidity of the nuclear substance

for
most dyes.
is
good example for practice
in double
staining
tion,
the hsematein- eosin combination.
The
secy
attached to the slide with albumin

is
fixative,
and
cleared,
passed
is
down through
the grades of alcohol to
30%, and
tion of
then placed for one or two minutes in a solu-
Mayer's haemalum.*
I
The
cc.
section
is
then washed
alcohol, With
*Mix
gram haematein
dissolved in
50
cc. of
90%
50 grams alum dissolved in 1000
water.
Cool,
settle,
and
filter

in water
till
OBJECTS.
5"
the stain
is
removed from the cytoplasm

alcohol.
and passed through 30%, 50%, and 70% this stage it is immersed for a few seconds
of eosin in
At
in a solution
70%
alcohol to stain the cytoplasm,
and
is
then passed rapidly up through
90% and 95%

The
alcohol
and
xylol to be
mounted
in balsam.
nuclei should
be dark blue and the cytoplasm reddish yellow.
Another good stain for the beginner

to
to use,
though not
make
up,
is
the Wright modification of Leishman's

single solution yields a
blood-stain,
which with a
most
beautiful differentiation of half a dozen cell elements.

Its use
is
described more fully in Chapter IX.
REFERENCES.
Chamberlain, C. J. Methods in Plant Histology. Chicago, 1901. Gage, S. H. The Microscope. Ithaca, N. Y., 1904. Lee, a. B. The Microtomists' Vade Mecum. Philadelphia, 1900. ZiMMEEMANN, A. Botanical Microtechnique. Humphrey, J. E.,
trans.
New
York, 1893.
CHAPTER
IV.
MICROMETRY AND THE CAMERA LUCmA.

I.
The Stage Micrometer.

whose spread
Objects
may be
directly
measured under the simple microscope by the use of

dividers,
is
determined, under the lens,
by means of a steel scale divided into fifths of a millimeter. With higher powers, some special standard is
required;
intervals of
and
.i
glass slides bearing fine lines

.01
ruled at
and
mm.
are prepared for this purpose.
These are known as stage micrometers.
The
imit in
is
which the measurement of microscopic objects
com-
monly expressed
is
.001
mm., known as the microletter

[t.
millimeter or micron,
and denoted by the Greek

is
Obviously such a scale

for
the only absolute essential
micrometry or measurement with the microscope,

it
and
would be possible
to find the
dimensions of an
slide
object
off the
by simply placing
it
on the ruled
and reading
is,
number
open
to
of spaces covered.
This process
Accurately
how-
ever,
two
objections.
graduated
stage micrometers, which are
expensive,
their
would be too
in this
it is
subject to breakage to
make
employment
manner economical.
Furthermore, on such a scale

52
MICROMETRY AND THE CAMERA LUCIDA.

difficult to
S3
measure small
lie
objects, since, as
mounted on
and
the slide, they tend to

their
obliquely across the scale,

its
edges would only rarely coincide with

rule, therefore,
is
lines.
As a
ment
2.
some
indirect
method
most
of measure-
adopted.
The
of
Ocular
Micrometer.
^The
satisfactory
method
This
measurement with the microscope involves
the use of another scale, called the ocular micrometer.

is
a circular piece of glass cut to
fit
in the eyepiece of lines
of the microscope
at
and bearing a
scale
made up
an equal, but not necessarily a known, distance

is
apart.
The micrometer
resting
to
be placed inside the eyepiece,

inside
it,
on the
sliding
diaphragm
which should image

lie
be so adjusted as
to lie in the plane of the real
formed by
the scale,
the objective.
Thus
the image will

it.
on
and may be measured by
The
value ob-
served
is
therefore the size of the virtual image of an
object formed by the objective in terms of an arbitrary

scale.
In order
to obtain actual
dimensions the ocular

to
micrometer must be standardized in order

the value of object of
its
determine
divisions as
size.
compared
this
to the
it is
image of an
simply nec-
known
For
purpose
essary to place a stage micrometer under the microscope
and focus upon
it
with the ocular micrometer in position.
The
ratio of ocular divisions to stage divisions (or rather
to the
images of stage
divisions)
can then be read
off
it is
and, knowing the actual value of the stage divisions,

easy to calculate the
number
of microns
on the stage
which correspond
to
one ocular division.

is
When
once
the ocular micrometer
standardized for a given system
54
of lenses

and length
of draw-tube, the stage micrometer
may be
discarded.
it
In using the ocular micrometer have seen, that the diaphragm,

at
is
necessary, as
we
in the eyepiece
should be
a proper level or the scale will not be
distinct.
Too
strong a light also blurs the image of the micrometer

divisions.
The
object
to
be measured and the scale
should be arranged in relation to each other, by rotating

the eyepiece
and adjusting the sUde,
so that one edge of
the object shall coincide with the edge of one of the lines
of the scale.
Since these lines have a considerable thickto refer the object to corre-
ness,
care
must be taken
sponding edges of the
lines at its extremities.
The
difh-
culty of estimating the boundaries of objects with accu-
racy
is
always great; and the limits of precision of meas-.

in the best microscopic
urement
3.
work
is
about
.2
/i.
Measurement with
the
Camera- Lucida.
Any
will
method by which
serve for
the image of an object
may be comand, although

satisfac-
pared with the image of a scale of known dimensions
measurement with the microscope

is
the ocular micrometer
the simplest
and most
tory process, there are several others, of which one involv-
ing the use of the camera lucida

tioned.
may be
briefly
menis
This apparatus, whose external appearance

in Fig. 26,
fits
shown
over the eyepiece of the microscope.
It contains
a system of mirrors which reflect the rays of

laid beside the microscope, so
light
coming from a paper
that they enter the eye along with the rays
which pass
up through the microscope.

object
Thus
the
image of the
upon the
stage
and
that of the paper are super-

MICROMETRY /IND THE CAMERA LUCIDA.
posed one upon- the other.
matically
the
Fig.
55
27 indicates diagram-
general arrangement of the mechanism.

is
At P, just over the eyepiece,
a cube
made up
of
two triangular prisms of
glass with a silvered
surface
between them pierced by a central opening.

this
Through
from
opening the rays from the microscope pass, while
those coming from the paper,
AB,
are reflected
first
the swinging mirror at the side
and then upward from
Fig. 26.
Camera Lucida.
(After Gage.)
the silvered surface of the prisms.

distortion
it
In order to avoid
is
necessary that the mirror should be at an
angle of 45" to the surface on which the paper rests.
In order
to
bring this about
it
is
necessary to sup-
port the paper on an inchned surface

the microscope extends outward
when
the base of
so as to interfere with
view of the
table.
The adjustment
of light with the

difficulty.
camera lucida
is
matter of considerable
If
the light passing
through the microscope be too
bright,- only the object will
S6

if,
be seen, and
the paper
invisible.
is
as
more commonly
occurs, the light
from
the strongest,
the object becomes almost

is
When
the illumination
properly balanced,
both object and paper should be clearly seen, and the

point of a pencil on the paper
outline
of
may be made
accuracy.
to trace the
this
the
object
with
Obviously
Fig. 27.
CouKSE or Rays
in
the Camera. Lucida.
(After Gage.)
device, primarily intended as
an aid
to the
making of
drawings of microscopic objects,

etry.
may be
used in microm-
With a given arrangement

drawing on the paper
will
of apparatus, the size

size of
of the
depend upon the
the object.
By removing
the latter
will
and
substituting a
stage micrometer
whose image
be superposed upon
that of the drawing, the size of the original object
may
be read
off directly.
MICROMETRY
4.
ySND THE
CAMERA
LUCIDA.
57
Determination of the Magnification of the Micro-
scope.
Îf
the image of a stage micrometer be projected

in pencil,
it
upon paper and drawn

possible,
will obviously
be
by comparing the actual
size of the
drawing
scale, to
with the
known dimensions
of the
micrometer
measure the magnification effected by the microscope.
The
is
size of the picture will,
however, vary with the
disit
tances between the paper and the camera lucida,
and
necessary to adopt for this distance a standard value,
generally 250
mm.
REFERENCES.
Carpenter, W. B., and Dallinger W. H. The Microscope and its Revelations. London, 1901. Gage, S. H. The Microscope. Ithaca, N. Y., 1904.
CHAPTER
V.
THE MICROSCOPY OF THE COMMON STARCHES.

I.
Origin and Nature of Starch.

for existence
ÂU
living
things
depend
upon the
and
potential energy of organic
compounds which they take

to simpler substances;
in as foods to
and break down

this constant
make good
drain upon the limited stock of organic matter in the

-world there
is
only one source of supply, the chlorophyll
bodies of green plants. Here rays of sunlight are absorbed,
and by
to
their energy
carbon dioxid and water are united

set free;
form
starch,
is
oxygen being
the kinetic energy
of the sun
thus transformed and stored

structure
up
of
in potential
form.
Ultimately the whole

this process, as
organic
life
depends on
rested
the universe, of mythology,
upon the back

is
of the fabled tortoise.
Starch
an insoluble carbohydrate substance belong-
ing to the cellulose group and having probably the formula

(CsHioOs)^.
It occurs as a white
powder made up
of
small grains a few hundredths, or rarely a tenth, of a milli-
meter in diameter, of a shape varying with the plant by
which
it
is
formed, but more or less characteristic for
each species.
The
grains are in general built
up
of con-
centric layers, the outer one resembling cellulose, while

5
MICROSCOPY OF THE COMMON STARCHES.
59
the inner layers contain an increasing proportion of water.
At the center there
is
frequently a small open space of

as the hilum.
much
less density,
known
Starch
may
be
seen in situ in the chlorophyll bodies of

process of active vegetation; but
it is
many
leaves in
rapidly changed to
is
sugar by the enzymes of the plant, and in that form
conveyed to the
root, the tuber, the stem, or

it
any other con-
venient tissue, where
may
be reconverted into starch
and stored
for future use.
The
seeds are particularly

roots
rich in such reserve material,
and with underground
and stems they furnish the principal sources

a commercial product.
2.
of starch as
Refining of Starch.
^The processes of starch manufrom the

tissue in
facture are in general simple, involving only the mechanical separation of the grains
which they
are embedded.
Thus, in the production of potato-starch,
the tubers after washing are ground

to
up
in a conaminutor
break the
cell
is
walls as far as possible.
The milky
fluid
produced
passed through a sieve with openings
about ^/bo of an inch in diameter which allow the grains

to pass while retaining a large proportion of the pulpy
tissue.
ties
still
The
starch
is
then separated from the impuri-
present, either
it
by
settling, in
tanks or in long
troughs through which

gal machines; finally
flows very slowly, or by centrifu-
it is
dried
and barreUed
for market.
fifth of
In potatoes the starch makes up perhaps a

total
the
weight and
four-fifths of the dry substance;

it is
but in
nitro-
the various grains
found in combination with

etc.,
genous compounds, glutens,

plicate the refining process.
which somewhat com-
Corn-starch
may
be sepa-
6o

is
rated with comparative ease, and

the starches;
the cheapest of
all
but in wheat and rice the union between

is
carbohydrates and albuminoid matter

introduce serious dif&culties.
essary to use strong alkalies;
so close as to
it is
In the
latter case
nec-
and with wheat the same and then allowing
end has generally been obtained by moisteniixg and thus

swelling the grain, bruising
it
it
to
stand and ferment until the nitrogenous substance has
been partly rotted and disintegrated.

feature of this
The
unfortunate
method
lies in
the necessary loss of the
glutinous constituents of the grain which possess high

nutritive value;
in certain processes the fermentation
is
omitted and gluten and bran are obtained as by-products.
The
terials.
cost of the various starches
depends both on the

of the
difficulty of separation
and on the value
raw malist
Its
range
is
indicated
by the following
of
prices compiled
by Dr. H. W. Wiley
for the year 1899:
MEAN WHOLESALE PRICE OF VARIOUS STARCHES.

Cents per pound.
Corn
Sago
flour
46-1 61
.
3-73-3 98
Potato
Tapioca
flour
4.21-4 57 4 63-4 96
.
Wheat
Rice
5 7
00-9 00
50-9 00
3.
Commercial Uses
of
Starch.
Wheat-starch
but
it
was
well
known
first
to the ancient
it
Greeks and Egyptians, having

said, in Chios,
been
produced,
is
was not

until the seventeenth century that the potato
6i
was used
as
a source of carbohydrate; and corn-starch

still
is
of course a
far the
more recent product.
These three are by
most important
of the starches.
As might be
inferred
is
from the
for
scale of prices given above, corn-starch
used
most purposes, the amount produced
in the United
States in 1903 being probably in the neighborhood of
75,000 tons.
Potato-starch ranks next with a produc-
tion of i6,coo tons in 1903,
and the amount
of wheat-
starch
was perhaps 10,000
tons.
The purposes for which this great supply is may be grouped- roughly under four heads:
used for a food material, for stiffening and
intended
starch
is
sizing, as
powder, or as a raw material for the manufacture of

other substances.
the
Mixed with nitrogenous

supreme importance
is
bodies,
in
form of
flour, its
;
of course as a
nutritive substance
and even the

no
purified product, in the
case of corn-starch, forms
insignificant contribution to
our dietary.
The
sago and tapioca flours are mainly
used for food, and the various arrowroots contain starches
recommended
as particularly desirable for invalids.
The
principal commercial importance of refined starch,
however, comes from the fact that when in contact with

hot water
its
grains swell
up and
burst, forming a thick
adhesive paste.
This
may
be best brought about by
mixing starch with cold water and slowly pouring the thick milky fluid into boiling water which is meanwhile
agitated
by constant
stirring.
The
name
paste thus formed
imparts to textile materials a high degree of lustre and

that stiffness from which the
of starch
(German,
62

Its adhesiveness Its stiffening
Starke) was originally derived.

it
makes
power
desirable for bookbinder's paste.

it
gives
supreme importance
in the laundry;
for society
has not yet outgrown what the Puritan divine described

as
"a
certaine kinde of liquide matter
which they
call
starch,
wherein the devill hath willed them to wash and
drie their rufHes,

stiffe
which when they be
dry, will then stand
and
inflexible
about their necks."
The
quality of
forming a
cotton-
size gives starch
an increasing importance in
and paper-mills, print-works and bleacheries.

stairch is
As a powder,
preparations,
in
used in
many
and
pharmaceutical
other
baking-powders
products
where some
finely divided neutral substance is desirable,
and
for
powdering the forms

starch
in printing-houses.
Finally,
when heated
in the dry condition to
i5o-2oo F. yields a soluble dextrin, sometimes called British
gum, extensively used

treated with
is
as a substitute for
gum
arable.
When
starch
weak
acids or with certain
enzymes
converted into dextrins and sugars and
may
thus
serve as the
raw material
for the
manufacture of glucose,
maltose,
4.
and ultimately
of alcohol.
Microscopical Examination of Starch.
^Since
the
chemical composition of the various starches

the
is
identical,
microscope offers the
only satisfactory" means of
studying them.
Fortunately the appearance of starchis
grains from the various groups of plants

teristic;
quite charac-
and one familiar with the commoner forms can
easily detect the sophistication of other substances, like
mustard, by means of starch, the presence of foreign

bodies
(minerals or seed-hulls) in starch, or the adul-
MICROSCOPY OF THE COMMON ST/IRCHES.

teration of
variety.
63
one kind of starch with another cheaper
Being
finely
powdered, starch
is
in a condition
admi-
rably adapted for microscopic examination, and needs
only to be mounted in some

characteristics
medium which
If
will set off its
by proper
solid
contrast.
examined dry, the
edges of the grains appear so black as to obscure the

view,
and most
so high
mounting media have on the other

of refraction that except with
hand
an index
polarized light the starch becomes too faint.
Water with
and
if
ordinary illumination gives a good picture;
the
hilum
is
to be particularly studied, a
like
medium
of higher
density
clove-oil
detection of starch
may may be
be useful.
Sometimes the
it
aided by staining
with a
dilute solution of iodine,
which produces the blue-black
color of the iodo-starch reaction.

5.
Potato-starch.
Potato-starch
in
is
produced in conin certain
siderable
amount both
States,
New
it
England and
Western
sizing of
and
is
used mainly in print-works for the

is
warp yarn before

particular
woven.
Other starches
make a more even and permanent

for
this
paste, as a rule, but
purpose
potato-starch
seems best
adapted.
As viewed with a hand-lens

that a
potato-starch
may
be at
once distinguished from most other varieties by the fact
mass of
it
appears to be studded with glittering
points, while corn-starch or wheat-starch is of a dull
dead
white.
Under
the microscope potato-starch
is
seen to be
made up
of large grains, .05-. 12
mm.
long, of a flattened
ovoid shape, with a smooth and regular outline.
As
in
64

there
is
all starches,
a considerable variation both in shape
and
size;
many
grains are ellipsoidal, soni, three-corspherical.
nered,
and the smaller ones almost
Near one
dot,
end of the grain the liilum appears as a well-marked

surrounded by rather faint concentric
ellipses
which form
an important
characteristic
of
potato-starch
and are
known
more
as the oyster-shell markings.
Sometimes two or
hila
appear in a single grain, and often groups of

to
two or three grains seem

Fig. 28 (5).
6.
grow
together, as
shown
in
Wheat-starch.
Wheat-starch
its cost, is
makes a very even
paste and, in spite of
used in
many
It is
processes
where especially
fine
work
is
necessary.
is
mixed with
the colors in printing cloth

ing, dyeing,
and
utilized in the bleachIt is also largely
and
finishing processes.
used in paper-mills.
Microscopically,
the
grains
of
wheat-starch
usually
appear as somewhat irregular
circles,
but when tipped up
on edge
their true shape is seen to
be lenticular.
In
size
it is
they vary from very minute points up to .04 mm., and
noticeable that the grains are mainly of two sizes, quite

large
(Fig.
and quite
28
(i)).
small, intermediate grades
being rarer
out
The hilum can sometimes be made
in the largest grains;

tric circles
under the same conditions concen-
may
indicate the natural layers of the grain,
though as a rule neither hilum nor markings are apparent.

7.
Corn-starch.
Corn-starch,
as
we have
seen,
is
in
America by
far the
most important of starches, being used
in the kitchen, the laundry,
and the
mill for diverse pur-
poses, as well as for the manufacture of dextrins
and

sugars.
It
65
may
often be found as an adulterant in the
more
costly starches,
and
in spices
and other
foods.
size.
Its grain is easily recognized,
being of
medium
''^ô:
o o
s>
(J
CO
(Q
^ o ^ ^6 Q
Fig. 28.
^The
1.
2.
Commoner Starches.
240 diameters.
(Redrawn,
4. 5.
6.
after Scliimper.)
Wheat-starch.
Corn-starch. Rice-starch.
3.
Tapioca-starch. Potato-starch. Bean-starch.
.0I-.02
mm., and
potato.
of a characteristic polyhedral form,

it
its
angularity at once distinguishing
from the starches
of
wheat and
Since
it is
not flattened like the
latter.
66-

approximately isodiametric,
it
h\xt
shows very black edges
in air-
and water- mounts.

is
Concentric layers are absent;
but the hilum
sharply marked,
showing in water-
mount
effect
the
form
of a cross with cracks radiating out
toward the periphery of the grain.

disappears and the hilum
is
In denser media
this
seen to be round (Fig.
28
(2)).
8. Rice-starch.
Rice-starch
It
is
resembles
that
of
corn
in a general way, being polyhedral in shape with distinct

facets
and
angles.
very
much
size.
smaller,
however,
having an average diameter of about .004-.008 mm., and

the grains are rather regular in
istic is
Another character-
found in the fact that the grains commonly occur
aggregated in masses.
(Fig. 28 (3)).
As a
is
rule
no hilum can be seen
This starch
used to some extent as an
fadulterant, and
9.
as a constituent of various powders.
The Starches
of the
Pea and Bean.
^The pea
and
and
bean, as well as certain other plants of the order Leguminosae, have starches of a very characteristic type;
though not of special importance in themselves, an acquaintance with their appearance

is
an important aid
in
the detection of adulteration with the ground-seeds of

these species.
Bean-starch, which
may
be taken as an
example of
this
group, has grains of an elliptical or kidsize of
ney shape with a

the
.02-.06
mm.
The hilum has
form
of a
slit
running the long way of the grain,
while distinct concentric layers
may
be made out in a
good
10.
light (Fig.
28
(6)).
The Arrowroots.^-Arrowroot-starches
and
are
exten-
sively u.sed as foods for invalids
for certain other
MICKOSCOPY OF THE COMMON STARCHES.

special purposes,
67
and are rather
liable to sophistication.
The
characters of the group vary widely, and
we can
only
consider one
common
its
example.
Bermuda arrowroot has

and the posses-
a starch which in
flattened ovoid shape
sion of oyster-sheU markings resembles that of the potato.
The
grains are however smaller, .02-.03
mm., and the

is
hilum, usually at the larger end of the grain,
typically
elongated to the form of a
slit.
The
smaller grains are
not rounded as in potato-starch, but have the same shape

as the larger ones.
II.
Sago and the Cassavas.
^Tapioca
is
derived from
the root of the bitter or poisonous cassava, the hydro-
cyanic acid which
it
contains being driven off in the pro-
cess of manufacture.
The
sweet cassava, in which this
substance
is
not present, yields a similar starch which
has recently been introduced into the trade for certain

sizing processes to
which
it
is
supposed to be specially
ciris
adapted.
Tapioca starch-grains are very smoothly
cular and often

central
show a truncated end.
The hilum
and
slit-like.
The
size is .01-.02
mm.
(Fig. 28 (4)).
The
grains of sago-starch have a similar hilum,
and
occasionally
show truncated ends.
They
are larger than
those of tapioca (.02-.05 mm.), and phenomenally irregular,
the roughly ellipsoidal form being generally distorted.
REFERENCES.
Galt, H.
The Microscopy
London, igoo.
of
the.
more commonly occurring

1904.
Starches.
Leach, A. E. Maurizio, a.
Food Inspection and Analysis. New York, Berlin, 1903. Getreide, Mehl und Brot.
ScHiMPER, A. F. W.
Anleitung zur mikroskopischen Untersuch-
68
ELEMENTS Oh APPLIED MICROSCOPY.

ung der vegetabilischen Nahrungs- und Genussmittel.
1900.
Jena,
VoGL, A.
E.
Die
wichtigsten
vegetabilischen
Nahrungs-
und
Genussmittel.
Wien
S.
u. Leipzig, 1899.
Wiley, H. W.
Cassava.
istry.
The Manufacture
U.
of Starch
from Potatoes and
Department
of Agriculture, Division of
Chem-
Bulletin No. 58.
Washington, 1900.
CHAPTER
VI.
FOODS AND DRUGS AND THEIR ADULTERANTS.

I.
Microscopical Examination of Foods and Drugs.
of
In the examination of foods and drugs for the detection of

adulterants the microscope
in
is
of very great value;
and
many
cases
it
furnishes the only satisfactory

says, in his treatise
method
analysis.
Mr. A. E. Leach
on "Food
Inspection and Analysis":
"The
chemical constants of
many
of the adulterants of coffee
and the
spices
do not
always differ sufficiently from those of the pure foods in
which they appear
to
be distinguished therefrom with
accuracy and confidence by a chemical analysis alone.
On
the other hand, one
who
is
familiar with the appear-
ance under the microscope of 'the pure foods, and of the

starches
ants,
and various ground substances used
as adulter-
can with certainty identify very minute quantities
of these materials,
when
present, with the
same ease
that
one can recognize megascopically the most familiar objects
about him."
starches,
The
already treated in Chapter V, furnish
perhaps the best examples of a case in which foreign

materials
present
may
be readily detected under the

of
microscope.
The
identification
such substances as
69
70
coffee

and mustard, containing a number
of
complex
tissues, is
more
difficult,
but a careful comparison with
samples of known purity insures reasonable certainty.
The
technique
of
the
microscopical examination
is
is
very simple.
The
substance to be observed
ground up
fine (so as to pass
through a 6o-mesh
fingers,
sieve), further com.-
minuted between the
mounted
in
water,
and
examined
directly.
It is
sometimes advisable to rub the

condition
powder
to
still
finer
by manipulating
the
cover-glass
and
slide
between the thumb and
finger.
The
process should not be carried too far, because in
tissues
which are broken up into very
fine
fragments the
Bet-
characteristic structures are often indistinguishable.

ter
views
of the
structure
of
opaque objects may be
obtained by clearing or rendering them transparent by

the
action
of strong alkalies,
caustic
this
soda, or chloral
is
hydrate.
necessary.
In practice, however,
not generally
The
and
student should be careful that whatever comes in

is
contact with the specimen to be examined

free
quite clean
from the contamination of previous specimens.
Several samples from different portions of the material
should be examined in order to gain an idea of
its
aver-
age composition.
tissue
Since the recognition of fragments of
depends largely upon the personal element, certain
points catching the eye of each observer, the comparison
with standard pure samples should never be dispensed

with.
2.
General
Nature
of
Food Adulterants.
A
is
very
thorough and systematic examination of foods
carried

on by the State Board
of
7^
Health of Massachusetts;
that
and
the
annual
as
to
reports of
board furnish valuof
able data
substances.
the
actual
condition
commercial
the
Spices,
coffee,
and
cocoa
are
most
analy-
important
sis is
foods
for
which the
microscopical
found available; while chocolate,

cassia,
tea, tobacco, all-
spice,
pepper, cayenne, paprika, cloves, ginger,
rnustard, nutmeg, vanilla,
cardamom, and numerous other

Cocoa
fre-
materials
may
be examined to advantage.
quently contains foreign cereals, starch, or sugar;

fee is adulterated with pea-hulls, peas,
cof-
chicory, wheat,
of the foods
and charcoal.
On
the other hand, in
some
advertised as substitutes for coffee a considerable admixture of coffee
may be
found.
Tea, cloves, pepper, and
mustard are most frequently adulterated with refuse portions
of the plant in question--tea-stems,. clov&-stems,
pepper-shells,
sionally very
and
mustard-hulls
respectively.
Occa-
bad samples
of cloves occur with a large
proportion of wheat, turmeric, and charcoal; of mustard,

nine-tenths wheat
olive-stones.
It
and turmeric;
of pepper, one- third
would be unprofitable, even
if it
were possible, for
the student to attempt to cover the whole field of microscopical analysis, since the detailed information involved
can best be acquired in practice when
it is
needed.
We -shall
their
therefore take
up only
three of the most im-
portant substances, coffee,
mustard, and pepper, with
commonest
adulterants, as types of the rest,
and as
illustrating the sort of characteristics
by which vegetable
food substances are identified.

72
ELEMENTS OF
3.
/tPPLIED MICROSCOPY.
of Coffee.
The Microscopic Structure

is
^The
coffee-
bean
the seed of Coffea arabica, a small tropical tree of
the family Rubiaceae, two of the semi-ellipsoidal beans

lying base to base in each of
its
berries.
The beans
are
brought into the market generally roasted, either in their

original
form or ground, and although adulteration

artificially
'
is
most easy in the second condition,
modelled
beans of foreign material are not unknown.
The
true
coffee-bean
is
made
of
thick-walled
cells,
approximately isodiametric, and packed with
finely
Fig. 29.
Microscopic STRtrcruRE of Coffee.

240 diameters.
(After
Schimper.)
granular
cells
material
containing
minute
oil-drops.
The
of the inner part are very characteristic,
showing
knotty thickenings of their walls, as indicated at

'Fig. 29.
in
More
peripheral cells (C) are smaller and lack
these swellings, while at the extreme outside of the

is
bean
the so-called silverskin, a thin glistening layer contain-
ing peculiar fusiform cells with wide walls pierced by

channels {A, Fig. 29).
rule, as large
73
The
first
two
tissues appear, as
opaque dark-brown masses, with the
cell
structure showing only at the edge.
The
detection
under the microscope of other
cell
structures than those mentioned at once serves to indicate the presence of adulterants.
Leach has pointed out
that even a naked-eye
examination sometimes reveals

chicory grains are apparent
foreign substances.
"The
from
their
dark and somewhat
gummy
and
of
appearance, and
can usually be recognized by crushing them between the

teeth.
Their
soft consistency
bitter taste are very
distinctive.
The
dull
is
surface
in
the
outside
of
the
crushed coffee-grains
marked
contrast to the pol-
ished appearance of the surface of the broken peas or
beans, often to be found as adulterants, while fragments

of
broken cereal grains are readily distinguished from

low-power magnifier, though perhaps not
coffee with a
easily identified
4.
by the eye alone."

other
Chicory and
Adulterants
is
of
Coffee.
^The
tissue
roasted root of Cichorium Intybus
one of the substances

coffee.
most commonly found mixed with

is
Its
presence
at once betrayed by the appearance of masses of
made up
of elongated cells of the type
stems of the higher plants.

walled and delicate, while the
large
common to the The outer layers are thinwoody tissue proper contains
fine
fusiform
cells
marked with
scar-like
cross-
hatching, the ladder-cells of the fibro-vascular bundles.
Occasionally, too, particles will be found which
show the
very peculiar branching, tubular structures of a homo-
geneous dark
color,
which are known as the milk-tubes.
74

general predominance of elongated
cells,
The
the presence
of
of ladder-cells
and milk-tubes, the absence
opaque
masses of dark-brown tissue and of granular cell-contents
make
the picture very different from that obtained in the
case of cofEee.
Various other roasted roots,

fruits
radishes,
carrots,
etc.,
such as
figs
and
pears, with grains of diverse sorts
are sometimes used as coffee substitutes.

setts the following
In Massachu-
have been found
Roasted peas, beans,
wheat, rye, oats, chicory, brown bread, pilot-bread, charcoal, red slate, bark,
and dried
pellets, the latter consist-
ing of ground peas, pea-hulls, and cereals, held together
with molasses.
cells,
In each case the presence of peculiar
other than the simple ones characteristic of the
coffee-bean, will reveal the sophistication.
The groundpea-hulls,
for
up
seeds of the Leguminosae, peas

aiê
and
example,
tected
quite
commonly
present,
and may be dep. 66,
by the starch-grains described on

from
and by
the outer layers of the hull, which

structure are
5.
their parallel
known
as the palisade
cells.

two
of Mustard.
is
^Mustard,
in
as
it
usually comes to the market,

of
a mixture of the
ground seeds
closely related herbaceous plants,
Brassica (Sinapis)
alba
and
nigra.
if
The
structure,
both cases,
is
very similar, and
the entire seed be ground
up and examined,
rent.
three distinct cell types will be appaflat
On the
outside are several layers of large

cells,
trans-
parent thin-walled
onal,
some rounded and some polygfaint
showing
in
glycerin
(c
concentric
markings
about a median area
and
d, Fig. 30).
These epider-

mal
layers
lies
75
are
difficult
to
them
layer
a characteristic
make out tissue known
clearly.
Within
as the coluijinar
made up
of prismatic cells which, as seen
under the
microscope, are small and polygonal with heavy walls
and small
pearance.
central lumens, giving the whole a dotted ap-
In white mustard
(B. alba) this tissue is yelit
lowish, while in
black mustard (B. nigra)
is
dark
brown, determining in each case the general color of the
Fig. 30.
^Microscopic Structure of Mustard. 240 diameters.
(After Schimper.)
seed
(b,
Fig.
30).
fairly
Finally,
the interior
cells,
is
a tissue of
medium-sized,
fine,
(a,
thick-walled
packed with a
gray-green, granular material containing oil-drops
Fig. 30).
6.
Adulterants of Mustard.In a good table-mustard
the amounts of epidermal and columnar tissue present

are very small, since the hulls are largely sifted out
;
the
76
ELEMENTS OF
cellit
bulk of the sample should be made up of the inner

contents.
is
is
If
an excess of the outer layers be present,
evident that mustard-hulls have been added;
and
this
one of the commonest adulterations to which

is
this sub-
stance
subjected.
Wheat- and
rice-starch are
also
found not uncommonly, and furnish conclusive evidence

of
sophistication.
Yellow aniline dyes are sometimes
used to give the proper color to such adulterated samples.
Turmeric, which
longa,
tard.
is
is
the
ground rhizome of Curcuma
also well
adapted for the adulteration of mus-
It
appears under the microscope in small amor-
phous, intensely yellow, pasty masses, which stain blue
with iodin, being

7.
made up
largely of curcuma-starch.
of Pepper.
Pepper
is
the pulverized seed of Piper nigrum, a shrub cultivated
mainly in the East Indian islands.

about
5
The
is
dried seed
is
millimeters in diameter
hull.
and
covered with a
brownish
If this hull is
ground up with the grain,

pepper.
we have the ordinary black made by macerating the fruit
White pepper
is
in water before drying,
and
detaching the hulls by friction.
In ground black pepper a number of
tissues
may
be
made
out,
since
the many-layered hulls are relatively

central
thick as
portion.
compared with the more homogeneous
The
latter
forms the bulk of the preparation,

It consists of
and
is
quite characteristic in appearance.
irregularly angular whitish masses, seen to be
made up
these
of polygonal cells
packed
Fig.
31).
full
of
very minute starch-
grains
(5
and
55,
With a high power

and
grains themselves appear polygonal
closely aggre-

gated into masses, while their size
is
77
very small, averaging
about .003
mm.
Besides these starch masses at least

of
cells
two characteristics kinds

appear in black pepper.
from the hull
may
so
The 'outermost dark-brown fragments made up of the

called, polygonal, thick-walled
cells
layer furnishes
stone-cells
with a small dark
lumen and
striations radiating out
from
it
(a,
Fig. 31).'
Fig. 31.
^Microscopic
Structure of Pepper.
240 diameters.
(After
Schimper.)
The
cells of the
inner part of the hull occur in lighter,
yellowish-brown masses, and have a somewhat similar

structure except that their
their walls thinner (&,
lumen
31.)
is
much
larger
and
Fig.
Smaller fragments of
irregular,
parenchyma
walled
of oil
cells
tissue
may be
seen, with
thin-
and occasional large spaces where deposits

fruit (c, Fig. 31).
were present in the
White pepper made up simply of the
central part of
the seed contains practically nothing but the irregular,

silvery
masses of pepper-starch with occasional small

tissues.
fragments of the outer
78
8.
ELEMENTS OF /IPPUED MICROSCOPY.

Adulterants of Pepper.
adulterated
of
Pepper
spices,
is
one of the most
commonly
and may be found
mixed with a wide variety of substances.

of mustard, the
As
in the case
ground hulls of the seed
itself
furnish a
frequent sophistication.
the United States
is
The most
it
general adulterant in
is
perhaps buckwheat-starch, which
not easy to detect since
closely resembles that of the
pepper in shape and occurrence.

are,
The
individual grains
however, about twice the
size of the pepper-starch,
and the masses,
as a rule, are also larger.

is
In Germany
dried and ground bread
often used, as well as the bark
of trees, bran, sawdust, pulverized nut-shells, of mustard, rape, peanut, linseed, or almonds.
and
hulls
In France
and
form
in this country
ground olive-stones are a
common
adulterant.
They
are
made up
principally of large fusi-
stone-cells
which resemble those of the pepper
except in their size and the fact that they are practically
colorless.
The
occurrence of any substance other than the
normal
starches
tissues of the pepper,
and
in particular of foreign,
characteris-
and the elongated
cells
and tracheids
tic of plant-stems, will suggest the presence of
some adul-
terant.
In
all
these instances only careful comparison with a
series of
known pure
substances will enable the analyst

is
to determine certainly which of these foreign materials
present.
79
REFERENCES.
Geeenish, H. a.
Drugs.
The
Microscopical Examination of Foods and
Philadelphia, 1903.
KoNiG,
J.
Die
menschlichen
NahrungsAnalysis.
und
Genussmittel-
Berlin, 1904.
Leach, A. E. Leach, A. E.
tion.
Food Inspection and
New
York, 1904.
Microscopical Examination of Foods for Adultera-
Thirty-second Annual Report of the State Board of
Health of Massachusetts for 1900.
MoELLER,
J.
Mikroskopie der Nahrungs- und Genussmittel aus

Berlin, 1886.
dem
Pflanzenreiche.
ScHiMPER, A. F.
W.
Anleitung zur mikroskopischen Untersuch-
ung der
igoo.
vegetabilischen Nahrungs-
und Genussmittel.
Jena,
ViLLEERS, A., et Collin, E.
Traitd des alterations et falsifications

Paris, 1900.
des substances aUmentaires.
TscHiRCH,
VoGL, A.
A., u.
Oesterle, O.
Anatomischer Atlas der PharmaLeipzig, 1900.
kognosie und Nahrungsmittelkunde.

E.
Die wichtigsten vegetabilischen Nahrungs- und

Berlin, 1899.
Genussmittel.
CHAPTER
VII.
THE EXAMINATION OF TEXTILE

I.
FIBRES.
The Kinds
the
of Textile
Fibres.
^The
is
word
fibre
is
derived from
or filament."
Latin
fibra,
and
signifies
"a thread
The most
primitive application of natural
fibrous materials to textile purposes
probably in the
use of grasses and osiers for weaving.

period, however, primitive
fibres of
cloth.
At a very early
man
learned to manipulate
a closer texture in the manufacture of coarse

long, fine hairs of plants
The
races
and animals among

attention;
many
must early have attracted
thus
the application of cotton
and wool dates back from be-
yond recorded
bast-layer
history.
In Egypt the use of the fibrous
from the stem of the flax-plant appears to have

According to Chinese
silkworm
antedated even that of cotton.
tradition, the application of the secretion of the
to textile purposes
was made by the

hemp,
first
empress of the
the prin-
nation.
To-day, four classes of fibres

flax,
cotton,
the
cipal vegetable hair;

all
jute, ramie,
and
sisal,
typical bast-fibires;
wool and a few
less well
known
of
animal hairs;
and
silk
still
make up
textiles
most importance.
The
leaves of certain plants furnish
filaments essentially similar to the bast-fibres mentioned

80
EXAMINATION OF TEXTILE
above.
FIBRES.
8i
The
leathery covering of the cocoanut suppHes

etc.
a fibre used for matting,
Asbestos illustrates the
possibility of using a mineral substance for textile pur-
poses.
Finally, various artificial fibres are
made from
is
metals and from cellulose derivatives.
In the
identifiLca,tion of textile fibres
the microscope
of prime importance.
Animal
of
fibres in general
may
by
be
the
distinguished
fact that
from those
vegetable
origin
both wool and
silk are soluble in
5%
caustic
soda, while the vegetable fibres,

are not thus affected.
of
made up
of
of cellulose,
Various color reactions are also

individual fibres,
value.
For the determination

is
however, the microscope
most
satisfactory, since
its
even
of the closely related bast-fibres each has

tic
characteris-
appearance.
Furthermore, the quality of different

is
samples of the same kind of fibre

size,
evidenced by the
twisting,
effect of
and external
structure of the filaments.
The
chemical reagents and the tensile strength

is
of individual fibres under various conditions
studied
with the aid of the microscope in

tories.
2.
modem
textile labora-
The
Cotton-fibre.
The
down
cotton-fibre
is
the vege-
table hair borne on the seed of plants of the genus Gos-
sypium, serving Hke the
of the thistle for the distri-
bution of the seeds by wind.

the
These plants belong
to
Mallow
to
family;
some are herbaceous and others

feet in height.
grow
be bushes twenty
As
the seedof
capsule opens a rich white boll bursts out,

the cottonseed with the fibres at
its
made up
free end.
The
cotton-plant grows well
between 45 north and
82
ELEMENTS OF /IPPLlED MICROSCOPY.

soil
35 south latitude, given proper

ply
of
and a uniform supIndia,
moisture.
The United
States,
Egypt,
China, and Brazil are the most important cotton-pro-
ducing nations in the order named; though

countries devote large areas to
its cultivation.
many
other
The most important

ton for the market
fibre
is
process in the preparation of cotginning, or the separation of the
from the seed-cotton, which as picked contains

its
two-thirds of
weight in seeds.
The modem
process,
evolved from that which Eli Whitney was largely instru-
mental in developing in 1794, consists in exposing the

cotton to the action of a series of fine-toothed circular
saws which tear
off
the fibre and carry
it
away through
the grid in which they revolve.
The
fibre, as
thus obtained,
is
a hollow ribbon (Fig.
32) spirally twisted at frequent intervals with the edges of the ribbon so sharply
marked
off
from the central canal
that they appear like swollen cords.

fibre
is
The
section of the
not,
however, dumbbell-shaped, as this might
suggest, but elliptical or
somewhat
crescentic, the
lumen
is
of the canal following the outer contour.
The
canal
narrow in American and Egyptian cotton, and broader in

that
from India.
it
By
following a single fibre carefully

it
along,
will
be noticed that
tapers to a
somewhat
is
blunt point at one end and at the other extremity
broken
off
sharply where
it
was attached
to the seed.
The
mm.,
length of the
cotton fibre varies from 20 to 40

/^.
its
diameter, from 10 to 20
is
Vj"
-j^
treated with a
The
cotton-fibre
mainly
cellulose,
|
covered with a fine
cuticula of different composition.
When

EXAMIN/ITION OF TEXTILE FiBRBS.
solution of hydrate of copper in
dissolves
83
ammonia, the
cellulose
and
swells
up
the cuticula into bubbles between
the twists of the fibre, while at the twists the cuticula contracts, giving the characteristic
beaded appearance shown
in Fig. 31,
e.
Much
of the cotton in textile goods has been subjected
Fig. 32.
The
Cotton-fibre.
(After Hassack.)
200 diameters.
to ,the process of mercerization, or
immersion of the cloth
in a stretched condition in strong caustic soda solution.
Cloth thus treated
is
stronger, takes dyes better,
and has a
closer, firmer texture
and a fine glossy appearance. and the

fibres
Under
the microscope the individual fibres appear as crinkly,

irregular cylinders instead of fiat ribbons
central
canal
is
much
shrunken.
Sometimes the
are so

84
swollen that even the twists cannot easily be seen in a
water-mount; they show more

3.
clearly,
however, in
of
air.
Bast-fibres in
General.
Most
the
vegetable
fibres in
commercial use are derived from the bast-layer
of the dicotyledonous flowering plants.
The bast,
more or
or, as it is called
by botanists, the phloem,

and since
it
is
it
a
is
fibrous layer lying just under the bark,

less
developed in
all
the higher plants,

textile fibres
would
be theoretically possible to obtain one of them.
from any
In preparing the
fibre
from those herba-
ceous plants actually in commercial use, the stems are

first
retted or allowed to ferment
under water so that the
gummy
substances which hold the tissues together
may
be dissolved.
Next
they- are scutched or exposed to the
action of beaters which break
up the outer and inner

adhering
friable tissues, leaving the elongated bast- cells
together in threads or bundles.

find
it
The
bast-fibre, as
we
in
commerce,
is
thus
made up
of a
group of
cells,
not of a single
cell like
the cotton-fibre.
When
bast-cell
further broken
up under the microscope, the

less
appears as a more or
elongated spindle
with a central canal where the living protoplasm once

lay, the cell wall of cellulose
only being
left.
Both ends
are symmetrically pointed.

section,
In
size,
proportions, crossof various
canal,
and markings the

distinguished.
bast-fibres
plants
may be
We
shall consider briefly
the six fibres which have most commercial importance

flax,
hemp, Manila hemp,
jute, ramie,
and
sisal.
4. Flax.
Flax
is
obtained from the stem of
Linum
usitatissimum, a
tall
herb widely cultivated in the central

regions of Europe
FIBRES.
85
and America.
The
seed of the plant
furnishes linseed-oil, and the bast yields flax to be spun

into linen.
The
cells,
flax-fibre like
others of
its
class, is
bundle of
ends
each a hollow cylinder tapering at both

Its
cells
(Fig.
33).
are
distinguished
by
their
Fig. 33.
^The Flax-itibre.
(After Hassack.)
200 diameters.
large size (25 to 30
mm.
long by .02
mm.
in diameter),
and by the
fact that they are swollen or knotted at fre-
quent intervals.
The
central canal
is
narrow and nearly

is
circular in section, while the cell itself
somewhat
flat-
tened.
Thus when a number
of cells are
examined under
the microscope some will appear

others, while the canal in all of
much broader than

is
them
narrow.
At the
end the
flax cell tapers gradually to a very

at
sharp point,
tip.
and the canal disappears
some distance from the

86

5.
Hemp.
The
is
true hemp-fibre, to be distinguished

Sisal
from Manila hemp and

sidered later,
hemp, which
will
be con-
derived from Cannabis sativa, an herb

as the flax-plant,
with
much
the
same range
grown most
a strong
extensively, perhaps, in Russia

fibre,
and
Italy.
is
It is
though
Ifess
pliable
than flax, and
used for cordage,
sail-making, and the manufacture of certain other textiles.
The Cannabis
plant also yields the drug hashish.
Fig. 34.
^The Hemp-pibre.
(After Hassack.)
200 diameters.
The
in
its
hemp-fibre
cell
very closely resembles that of flax
microscopic appearance, being of the same size and
general shape
It
and showing the same knots and
swellings.
First,
may be
distinguished by two characteristics.

cell, is elliptical
its
the canal, like the
in cross-section, so that
distin-
while the
cell
on
narrow surface cannot be
guished from
flax, its
FIBRES.
87
broad diameter shows the broad

Second, the ends of the
cell are
canal quite distinctly.
comparatively blunt, and the canal runs up to the very

tip (Fig. 34).
6.
Jute.
The
jute-plant, Corchorus capsularis,
is
an
annual, native to the East Indies, and furnishes a long
smooth
fibre which,
however, quickly softens and breaks
Fig. 3S.
^The Jute-fibre.
It is
(After Hassack.)
200 diameters.
when
wetted.
used for making rope, coarse twine,

for the backing of various other tex-
and gunny-bags, and

tile
materials.
The cells of which the long jute-fibre is made up are much shorter than those of flax and hemp, being only about 2 mm. in length, while their width is nearly the
same as
that of the other fibres.
In-
any given
field of
the microscope numerous ends of
cells will
be apparent,
88

cells of
while with the long

are hard to find.
Linum and Cannabis
the tips
The most marked

ever, its canal,
peculiarity of the
j'ute cell is,
how-
which varies markedly
in diameter, at one
point occupying a large part of the
cell
and then shrink-
ing to an almost invisible line (Fig. 35).

7.
Ramie.
^Ramie or China grass

Islands.
is
the product of a
to China, Japan,
low shrub, Bcehmeria nivea, indigenous
and the Philippine
Its fibre is long
and
lustrous,
Fig.
36.^Ramie.
(After Hassack.)
200 diameters.
stronger probably than any other fibre;
it
is
used for
making
cells of
sail-cloth
and
for other special purposes.
The
their
ramie (Fig. 36) are easily distinguished by their
great length, sometimes reaching 50 cm.,

flat
and by
they
ribbon-like
structure.
In
cross-section
are
EX/iMINATION OF TEXTILE FIBRES.

pointed
of the
ellipses,
89
.03-.06X.01-.02 mm., with a wide canal

as the
cell.
same shape
Under
the microscope,
like
therefore, as one sees the
two views, ramie appears
mixture of a broad fibre with a broad canal and a narrow

fibre
8.
with a narrow canal.
Manila Hemp.
Bast-fibres are not

leaf-fibres are of
confined to the
fibro-
stems of plants, but are continued upward in the
vascular bundles which form the veins of the leaves.
In many plants these
such a character
as to be valuable for textile purposes, notably in the case of
Manila hemp and

Manila hemp
is
Sisal
hemp.
obtained from the leaves of
Musa
iextilis,
a palm native to the Philippine Islands and
North Borneo.
though more
for
The
fibre
is
extracted
is
by hand, and,
extensively used
brittle
than true hemp,
marine cordage.
The
cells
are distinguished from
those of Cannabis by their lesser length (about 6
mm.)
canal
and by
their smoothness, the knots,
though present, being
much
is
less
marked than
in the hemp-fibres.
The
wide and easily discerned.

9. Sisal
Hemp.
Sisal
hemp
is
obtained
from the
leaves of a cactus,
Agave
rigida, extensively cultivated in

It is a
Mexico, Central America, and the West Indies.
long coarse white fibre used for cordage and the manufacture of rough sackcloth,
hammocks,
all
etc.
The
by
cells
are distinguished from
other bast-fibres
are 1.5-4
their
smoothness and regularity.
They
mm.
long and 20-32 n wide, approximately cylindrical, with a wide and well-marked canal, and tapering to a fine point
at the end.
As
in other cases,
comparison with material
9
of
known composition
furnishes the only sure criterion
for identification.
Ânimal hairs are 10. Wool and Other Animal Hairs. much more complex structures than those of plants, being
not only multicellular, but composed of several distinct

layers of cells.
'
In the growth of a hair the epidermis or

is first
outer tissue of the skin
folded in to form a minute

the base of this follicle
pit called the hair- follicle.
From
the hair grows out,
its cells
dividing off from the epideris
mal
tissue below.
The
hair itself
is
made up
of three
distinct
zones.
cells
In the center
an axis of irregularly
Outside
is
rounded
this,
known
as the medullary layer.

of the hair,
and forming the main portion

which
consists
the corcells;
tex,
of
elongated
spindle-shaped
and
at the periphery is a cuticular layer of flat overlapping
scales covering the hair like the shingles
on a
roof.
In
to
the cortical cells the hair

its color,
is
situated the pigment
which gives
and
to the disintegration of these cells

is
and the formation
of air-spaces in their place
due the
whitening of the hair in old age.

Externally the principal characteristic of
hair
is
the
animal
the presence of the scales of the cuticle
which give
the fibre the effect of being
marked with
at its
fine transverse,
anastomosing
as
fine
lines
and which
edge (Fig. 37) appear

cells
serrations.
The medullary
may
just
be
made
tions.
out beneath this cuticle as fine longitudinal stria-
The
',
general term wool
is
applied to hairs which are

will
spirally twisted so that
when woven they
hold
to-
gether to form a strong fabric.
In such hairs,
too, the..

scales are large
FIBRES.
91
and project prominently from the
fibre, so in-
that they interlock with each other
and materially
crease the compactness of the tissue.

varieties of sheep
Some
thirty-two
(bebnging
to the
genus Ovis) furnish
the wool of
commerce besides the llama, or alpaca goat, the Thibet or Cashmere goat, and the Angora goat, from
Fig. 37.
^Wool-fibres.
(After Hassack.)
200 diameters.
which mohair
is
derived'.
its
The
length of wool varies

fx;
from
2 to
20 cm., and
diameter from 10 to 100
the
larger fibres are
combed and spun
into worsted yarns,
the shorter are carded and spun into woolen yarns.
The
separation of wool from vegetable fibres under

is sufficiently
the lens
simple;
but the application of
the'
microscope only begins here.

animals
should
The
hair from various"
be
carefully
studied;
comparison
of
92
ELEMENTS OF y4PPUED MICROSCOPY.
sheep's wool with camel's hair will prove instructive,
showing the
lesser length of the latter, its
freedom from
twisting, the scales lying almost fiat against the surface,
and the granular spots

of
in the medulla.
Different grades
wool
may
then be studied, contrasting the high-grade

/i
merino wools with a diameter of 15
and 10 or more
wool of poor
twists in every centimeter of length with a
quality having perhaps four or five times that diameter
and not more than one

two
qualities, fineness
twist in a centimeter.
These
und
convolution, principally deter-
mine the quality

projection of
its
of wool, although its regularity
and the
scaly covering are also of importance.
sample of wool shoddy
may
profitably be examined;
various foreign fibres, dyed fibres, and torn and broken

fibres will
be apparent.
II. Silk.
One
The
other aniiftal fibre of quite a different
character remains to be considered, the secretion of the
silkworm.
larvae of
many moths
spin their cocoons
out of threads long and strong enough to be used for

textile purposes,
but that of
Bombyx mori
is
cultivated
most extensively for commercial purposes in

Japan, India, France, and Italy.
China,
The
of the in the
fibres
substance of which silk
is
composed
is
poured out
in a liquid condition
from two glands
at the anterior
air at
end
sets
worm, and when discharged in the

form
of solid
structureless
rods.
once
The
pair of
formed by the two large spinnerets are cemented
together by an incomplete cuticula of ent composition produced
somewhat
set of
differ-
by another
glands and
Joaown as seridn.
Under
the microscope such a double

thread at
first
FIBRES.
93
sight appears
Uke a broad
fibre
with a
central canal (Fig. 38, a).

ket,
In preparing
silk for the
mar-
however, the cocoons, before unwinding, are placed
in hot water to kill the chrysalis
and
to melt the
guramy
In
this
material which cements
all
the threads together.
process the single fibres become separated and each ap-
pears only as a semi-transparent cylinder (Fig. 38,
b)
with
Fig. 38.
Silk.
(After Hassack.)
200 diameters.
no
internal structure,
and no external markings except very

as the fresh secretion
fine
sets.
longitudinal cracks formed
Flecks of the ruptured cuticula and particles of
foreign material which
come
in contact with
liquid sometimes adhere

varies considerably
rate
at
to
the fibres.
it when semiThe diameter
from point
to point, according to the

its
which the worm was producing

jx
secretion.
Perhaps 10-20
would be a
iair average.
Ends, except
94

accidental breakage, are, of course,
when produced by
meters of silk
12.
is
absent, since the whole cocoon containing perhaps 500
often
made up
of a single piece.
Analysis of Fabrics.
In
the
examination of a
sq.
fabric a
sample should be taken 2-3
cm. in
size,
or
large
enough
to include all the different yarns
employed
in the pattern.
The warp and Ming

sometimes there
threads are then
separated into their constituents, and one of each kind
taken for analysis;

different
may
be a dozen
of the
is
yams
to examine.
Under a low power
microscope
(fifty
diameters) the nature of the fibres
determined in the different yams, taken in the proportion

in
which they occur in the
fabric;
a rough quantitative
analysis
may
thus be made.
REFERENCES.
Bottler.
Bottler.
Die animalischen Fasefstoffe. Leipzig, 1902. Die vegetabilischen Faserstoffe. Leipzig, 1900. Bowman, F. H. The Structure of the Wool Fibre. Manchester,
1885.
Brooks, C. P.
Cotton.
New York,
1898.
Dodge.
1897.
Descriptive Catalogue of the Useful Fibre Plants of the
World.
Report No. 9 of the U.

I.
S.
Department
of Agriculture
Hannan, W.
Hassack,
C.
The Textile Fibres of Commerce. London, 1902 Wodurch unterscheiden sich die Textilfasern ?
Leipzig, 1899.
Matthews,
J.
M.
The
Textile Fibres
New
York, 1904.
ViGNON, L. La Sole. Paris, 1890. Wilkinson, F. The Study of the Cotton Plant.
New York,
1899
CHAPTER
Vin.
THE MICROSCOPY OF PAPER.

1.
Paper and Paper-making.
The
A real
era.
earliest
written
records were probably

stones, leaves, bits of
made on such
of clay.
natural objects as
bone or wood,
or, as in
Babylonia
and Assyria, upon blocks

the celebrated papyrus,
paper, however,
in
was manufactured
Egypt many
thin, trans-
hundred years before the Christian
The
parent layers of tissue which surround the stem of the
papyrus plant were 'separated with some sharp
instru-
ment, superposed under water and then pressed and

dried.
Large quantities
of this product
were exported
from Alexandria
to all parts of
Europe and Asia, coming
into competition with parchment, prepared
by scraping
and
drying the skin of the sheep

process of
and
goat.
The modem
of cellulose
making paper,
as a thin layer
re-
derived from fibrous vegetable material

first
duced
to
a pulp in water, was
discovered by the
Chinese and introduced into Europe by the Arabs in the

eleventh century.
Cotton was the
first
material used
for this purpose, but
an Arabic manuscript on linen paper

is
still
bearing the date iioo
in existence.
The manuReaumur,
95
facture of paper from wood-pulp appears to have been

first
suggested
by the French
naturalist,
in
gS
1 7 19,
ELEMENTS OF yIPPLlED MICROSCOPY.
who
first
derived his idea from a study of the

their nests
way
in
which wasps construct
from
is
this material.
The
step in
paper-making
the treatment of the
raw stock with some chemical

dissolve the cementing
in order to
break
it
up, to
gums, and to separate the
cellulose,
which
is
always the principal constituent of paper, in as
pure a form as possible.
The crude
material
is
generally
boiled with strong alkali under pressure,

in a- tank
and then washed

is
from which the waste water
removed by a
acid
revolving drum.
Wood
paper
is
also
made by an
process, the stock being treated with the bisulphites of
lime and magnesia;
and mechanical wood-pulp
for a
low grade of paper
is
prepared by simple attrition against
stone surfaces without chemical action.
After treatment in the boilers and washers, the paperstock

stuff."
is
bleached, and at this stage

It is
is
known
as "half-
next passed through the beaters, in which a

it
wheel bearing knives breaks

condition.
up
into
a fine fibrous
auxiliary
Sizing, loading, coloring,
and other
processes are accomplished in the beaters.

fully
it
Finally, the
prepared pulp passes to the paper-machines, where

spread in a thin sheet of running water over a
belt of wire cloth.
is
moving endless
The
fibrous material
deposited in a fine and even layer passes on through

felted rollers,
which press out the
last of the
water and
compact
2.
its
texture.
The
Raw
Materials
of
Paper.
Obviously
paper
might be made from any material which can be ground
up
as
to a fine fibrous pulp;
and such bizarre substances

com-husks, cabbage-
seaweeds,
shavings,
sawdust,
MICROSCOPY OF PAPER.
97
stumps, and leather-cuttings have been used experimentally for this
purpose.
Cotton and linen rags

of
still
furof
nish
the
best
grade
is
paper.
The manufacture
Manila paper
an important industry.
Straw and
wood-pulp, however, furnish by far the largest yield of

coarse paper and cardboard.
the different materials
is
The
relative
importance of
table
indicated
by the following
from the United- States Census:

Paper-stock Manufactured in the U.
S.
in igoo.
Wood, cords
Wood-fibre, tons
Straw, tons
1,986,310
644,006
367,305
Old paper, tons

Rags, tons
356,193 234,514
99)3!
of
Manila, tons
3.
The Microscopic Examination
Paper.
Before
it
examining a sample of paper under the microscope,
should be torn into small bits and boiled in a one-percent

solution of caustic soda.
The wet pulp produced

fibres is
is
washed on a
fine sieve
and broken up by shaking

paper
in water.
The
with
identification of
by no means an
will
easy task, and requires careful study and comparison
known
substances.
The
student
be
much
aided by an admirable monograph prepared by Professor
W. R. Whitney and Mr.

lished in the
A. G.
Woodman, and pubfor September, 1902.
Technology Quarterly
The
authors suggest the following points to be noted in
the systematic
study of each specimen: size of
cells,
shape, length, width as compared to length, shape of

the ends, presence of knots or joints, whether the major-
98
ELEMENTS OP
/iPPLIED MICROSCOPY.
or bent or curled, markings,
if
ity of the cells are straight
size
and character
of the central canal,
any.
Polarized
light is often of great service in the identification of

fibres.
4.
paper
Analytical
Key
to
Paper
Fibres.
^The
In
authors
above mentioned suggest the following useful key for

the the
preliminary examination of paper.
this
table
word
fibre is restricted to those cells
which are very
long in proportion to their breadth.
ANALYTICAL SCHEME.
A.
Fibres are characteristic; other characteristic forms absent.
I.
Fibres are long; greater than diameter of field (Mag.

(i)
= 60).
Fibre has
many
is fine;
joints, knots, or projections,, especially

light,
by polarized
(a)
Fibre
quite smooth; joints not very notice-
able.
Paper mulberry
(5)
Manila hsmp Agave.

and promi-
Fibre
is
coarse; uneven; joints are large
nent.
Linen
(c)
Jute Hemp {Cannabis saliva),

dotted, circular, square,
Fibre shows overlapping scales.
Wool.
(2) Fibre has peculiar markings;
net-like.
(a)
Markings resemble
Spruce
(fir,
circular or square perforations.
Redwood
(6)
Red cedar Arbor
hemlock, tamarack, balsam)

vitce
Pine Cypress.
stem)
Markings
net-like, "feather-stitch," spiral.
Redwood
Fibre
(a)
is
Cypress Banana
regular.
(Jruit-
Ramie.
(3)
smooth and
fibres
Many
Cotton
(b)
Banana
resemble twisted ribbons.

{stalk
and
leaf-stem).
Fibres are round, cylindrical.

Silk
Sisal hemp Bark
0} cotton'Stalk.
II.
99
Fibres are short; less th^n diameter of field (Mag. =60).

(i)
Ends
of fibres frayed
Cotton rag
(2)
Mechanical wood
and
torn.
(conifers).
Ends
of fibres not torn.
Coir
Elm Willow.
-
Characteristic forms other than fibres are also present.

I.
Fibres are long; equal to or greater than diameter of
field
(Mag. =60).
(i)
Fibres quite broad in middle; tapering to point like a

bayonet,
(o)
One
or both ends of cells
drawn
out,
sometimes
terminating in a
Poplar
(6)
BirchPorpor gum.
walnut
tail.
Ends
Cottonwood
WhitewoodBlack Chestnut.
diameter of
field.
of cells cut off obliquely at blunt angle.
Holly
(2)
Fibres not varying greatly in width, ends needle-like.

(a)
Characteristic forms (cells) large;
covering i to ^
Bamboo
(b)
Sorghum bagasseRaffia Tulip.

cellular,
Characteristic forms small,
serrated,
or
pointed.
Straw
II.
Esparto Sugar-cane bagasseLive Oak.

field
Fibres are short; less than diameter of

(i)
(Mag. =60).
quite sharply
Cells comparatively long
and narrow;
buckeye.
pointed at one or both ends.
Magnolia
(2) Cells
TulipSweet
stubby;
fine,
prismatic,
ends
blunt
and cut
off
obliquely,
(o)
Quantities of short,
transparent material with
square ends.
Holly
(6)
Chestnut.
and
characteristic
of
Field fairly clear except for fibres
Pawpaw
(3)
Tree Willow. cherry Elm.
elements.
heaven
Maple Black
Cells fragmentary, broken; fibres short.
Groundwood

100
With a few of the 5. The Commoner Paper Fibres. most important materials of paper the student should
thoroughly famiUarize
linen,
himself.
jute,
The
silk
fibres
of
cotton,
hemp, manila,
and
have already been
treated with
some
fulness in Chapter VII.
Of
pine,
the
woods
dis-
used in the manufacture of paper pulp there are two

tinct
types.
The
Coniferas
spruce,
fir,
etc.
ex-
hibit cells of
markedly
different structure
from the Angio-
sperms, of which poplar and birch are the most important
examples.
Finally,
straw and esparto grass are
distinguished
6.
by a
third characteristic type of tissue.

of
The Structure
the
fir,
Gymnosperms.
^Wood-pulp
is
made from
the spruce,
balsam, larch, or hemlock
Fig. 39.
^Tracheid of a Conifee.
(After Herzberg.)
240 diameters.
mainly made up of the
cells
from the tracheids or

In
all
fibro-
vascular bundles of the stem.
the trees of this
group the structure
is
essentially the same.
The
-
cells
are long, exceeding the diameter of the low-power field,
and
are
of considerable breadth
(see
Fig.
39).
The ends
Sharp
often
contracted to
a rather acute point.
twists are
sometimes present, due to harsh treatment in
the preparation of the fibre.

of these cells, however,
is
The
characteristic feature
the presence of
numerous round

lot
or oval scar-like pits arranged in regular rows over their

surface.
In the case of pine,
lattice-like areas
with ob-
long openings appear at intervals; long

ings
7. cells
is
but the presence of
mainly of a single type and with discoid mark-
the characteristic of the Conifers in general.

of
The Structure
the
Angiosperms.
Pulp
made
broad
from the
commoner Angiospermous
trees
shows two
distinct elements,
long narrow fibres and short
Fig. 40.
^Tracheid of Birch.
(After Herzberg.)
240 diameters.
cells
with
characteristic
markings.
The two
are
species
most in use
for
cheap grades of paper
poplar
and
birch;
the longer fibres are very similar in both,

central
having
canal
of
variable
width and ends

to a point.
sometimes rounded and sometimes tapering
The more
thin- walled of these fibres often
show rounded
and oval pores penetrating the wall.
Smaller fibres with

swellings at intervals along their course are also present
in birchwood.
The
are
small,
broad
cells
of the fibrovascular bundles
more
characteristic.
In birch they are very numer-
A
Fig. 41.
B
Cells of Steaw.
(After Herzberg.)
C
240 diameters.
ous,
and are dotted with minute pores arranged
in
rows
at right angles to the long axis of the cell (Fig. 40).
The
At
distribution
of these
pores
is
somewhat
irregular.
the ends of the cell the cross-hatched transverse walls are sometimes seen.
In poplar the
cells of this
type are
less
103
closely packed.
numerous and show larger pores more

cells is often
At the ends of the
a long,
is
tail-like point,
and
the grating seen in the case of birch

8.
absent.
The Fibres
of
Straw and Esparto.
Paper
largely
from the stems
of the grasses
and grains
is
made made
con-
up
of long slender bast-fibres knotted or thickened at
regular intervals (Fig. 41, a).

stricted at these points
The
central canal
is
and
fine pores are present, passing
through the wall.

tissues are,
The
characteristic structures of such

cells
however, the
from the epidermis,
flat
and
of
somewhat variable
c).
length, with thick walls
and
serrated edges (Fig. 41,
In straw pulp a third type of
cell is present,
derived
oval,
from the internal pith

thin- walled cells with
layer.
These are
large,
rounded ends
(Fig. 41, J).
Esparto
or alfalfa grass (Stipa tenacissima)
may
be distinguished
from straw by the absence
of these pith cells,
and by the
smaller size of the bast and epidermal elements.
REFERENCES.
Cross, C.
F.,
and Bevan, E.
J.
Text-book of Paper-making.
London, 1900. Herzberg, W. Papierpriifung. Berlin, 1902. Whitney, W. R., and Woodman, A. G. The Microscopic Examination of Paper Fibres, Technology Quarterly, XV, 1902, 272.
CHAPTER
IX.
THE MICROSCOPE IN MJEDICINE AND SANITATION.

I.
The Microscope
in Biology.
^The microscope bears

that
to biology
much
the
same
relation
the
balance
bears to quantitative chemistry.
It is the
fundamental
instrument without which a true science of living things
would be almost impossible.
The
invention of the
of
compound microscope and the
construction
simple microscopes of high power at
the middle of the seventeenth century stimulated a dozen
observers to the study of the finer structures of plants
and animals.
Leeuwenhoek discovered
in
water and
in decomposing organic matter a teeming world of minute
forms of
life
invisible
to
the naked eye.
Hooke, the
structure
of
English botanist,
made
the
out
the
cellular
plants in his examination of cork.
Malpighi and Swam-
merdam
It
figured
microscopic details of the insect

success.
body with marvellous patience and

was
only,
however, after the perfection of the
achromatic objective about 1835 that these early labors

could bear abundant
fruit.
In 1838-39 Schleiden and
Schwann developed
their
that great generalization

all
which bears
104
name, the doctrine that
plants
and animals are
THE MICROSCOPE IN MEDICINE AND SANITATION.

built
105
up
of minute individual masses called
cells.
Quickly
the essential identity of the substance forming the cells
was postulated.
The microscope
thus grounded anatomy
upon
the firm basis of the cell theory;
and to-day
cytology,
the study of the minuter structure of the cell
itself,
forms
an independent subject
of
ever-increasing
cell is
importance.
Next
it
was discovered that the
not only the unit

living
of adult structure, but the
form
in
which every
organism originates from

the course of development
its
parent.
In the study of
single
cell
is
by which the
transformed into the mature organism, carried out by
Van Beneden, and their compeers, embryology came into being. Upon the anatomical and
Kolliker,
Hertwig,
embryological unity thus demonstrated under the microscope,
the
doctrine of
evolution,
the
most important
century,
scientific
contribution of
the
nineteenth
was
largely founded.
2.
The Microscope
in Medicine
and Sanitary Science.
Of all the branches of biology, none owes more to the compound microscope than the study of disease. The
conception of the body as a complex of protoplasmic
cells,
whose normal cooperation was the condition

easily
to
for
good health, led

might
arise
cell.
the
conclusion that disease

of
from the deranged functioning

Cellular pathology,
the
in-
dividual
associated
with
the
name
in the
arise
of Virchow, led along this Hne to great advances
knowledge of those diseased conditions which

activity of
from the abnormal
the
living
proto-
plasm.
Another group of maladies due
to the
invasion of
io6
parasitic
fevers,

plants
and animals, the
epidemic
plagues,
after
and
pestilences,
was comprehended only

by Leeuwenhoek
their exciting causes
were discovered by the microscope.

in 1680,
The
first
bacteria, observed
were
carefully
studied by Ehrenberg in the wonderful
decade of progress which followed the perfection of the

achromatic objective.
Twenty
years later Pasteur estab-
lished the casual relation of these
minute fungi
to disease;
and
in the next quarter of a century the
organisms caus-
ing anthrax, tuberculosis, fyphoid fever, diphtheria,
and
cholera were discovered.
These bacterial parasites are

but the microscope
causing
studied largely by culture methods;

is
also
indispensable.
The animal organism

artificial
malaria cannot be grown on
media,
and our
In
whole knowledge of
the
first
it
depends on optical methods.
few years of the twentieth century, the
brilliant
researches which revealed the parasites associated with
smallpox and scarlet fever were carried out with the

microscope alone.
The
researches into pure science which have led, and
are leading, to such advances in pathology
and
parasi-
tology cannot, of course, be treated here in detail.
Cer-
tain applications of the microscope to the study of the
body
fluids
and
of food material have, however,
become
in-
a routine part of clinical diagnosis and the sanitary

spection service.
cal
These technical applications
of medi-
microscopy come legitimately within the scope of our
work;
and a few
typical
methods for the
study of
pathologic conditions and of parasitic invaders will
now
be briefly considered.

3.
107
The Examination
of Urine.
Many organic diseases

etc.,
affect materially the

Its
composition of the urinary secretion.

specific gravity are significant,
amount, reaction, and
and chemical
value.
tests for sugar,
albumin,
are of
much
The
is
presence
of
certain, important
organized
elements
determined under the microscope.
The
precipitate
first
which forms
in
any urine on standing

better,;
must be
concentrated by sedimentation, or
by the use
of the centrifuge, transferred
by a
pipette to a
clean slide, covered, and examined with the high power.
The
edges of the preparation, where evaporation
is
taking
forrri
place, should be avoided, since the crystals which
here are not characteristic.
The
principal
objects
which
may
be found in an
examination of urinary sediments are crystals of certain

products of metabolism, red and white blood- cells, epithelial cells,
and
tube-casts.
The
inorganic salts precipi-
tated
may
be diverse in character, including uric acid
(clusters of
rhombic prisms and whetstone forms), acid
urates
ing),
(amorphous, granular masses soluble on warmcalcium oxalate (small octahedral crystals whose
diagonal planes look like the edges on the back of an

envelope),
and ammonium-magnesium phosphate
(long
prisms with bevelled edges, known as the cofSn-shaped

crystals).
(See Fig. 42.)
In hyperemia, acute nephritis,
and some other

in
conditions, red blood- corpuscles appear

ji
the urine as homogeneous, yelloAvish discs 7.5
in
diameter, sometimes distorted to a globular form or shrivelled
and crenated almost beyond

is
recognition.
cells
An
arbi-
trary line
drawn between "normal"
retaining

io8

and abnormal
colorless
their red color
and crenated
cells.
The
other formed elements of the blood, the white cor-
puscles (polymorphonuclear leucocytes), are larger bodies,
more or
less spherical
and possessing
several nuclei;
the
Fig. 42.
cells.)
urates,
Urinary Sediment in Catarrh of the Bladder. (Acid ammonium-magnesium phosphate, leucocytes and epithelial
(After
Ultzmann and Hofmann.)
200 diameters.
latter
may
be brought out clearly and differentiated

epithelium by allowing dilute
from the
cells of the renal
acetic acid to run
under the cover-slip when the nuclei of
the two types of cell
may be made out.
The
characteristics
THE MICROSCOPE IN MEDICINE AND SANITATION. 109

of the white corpuscles will be treated in the next section;
somewhat more
fully
is
but their presence in urine
of
much significance since it is these cells which mainly make up the whitish pus discharged from inflamed
surfaces.
When
the walls of the genito-urinary tract are

cells
seriously
affected,
detached from
its
lining
epi-
thelium will be noted in the urine.

of
The appearance
in
some
of
these
elements
is
indicated
Fig.
42.
de-'
Finally,
most renal diseases are characterized by the
posit in the fine tubules of the kidney of
an albuminous
substance which coagulates to form minute twisted tubes

(Fig. 43).
These
casts
found
in the urine
may
be clear
and hyaline or may be rendered granular by

of disintegrated epithelial cells or
the presence
may
contain undecom-
posed epithelial
cells or
pus
cells
or fat globules or bacteria
according to the special pathologic condition

they
arise.
from which
So-called
waxy
casts are denser
and more
an
sharply marked than any other type and
indicate
advanced stage
4.
of renal disease.
Examination of Blood.
plasma containing
^The
blood
is
a colorless
types
fluid or
cells of several distinct
whose number and
relative
proportions vary in
many
physiologic and pathologic states.
Few
clinical tests are
of more value to the physician both in diagnosis and
prognosis than the examination of this body fluid under

the microscope.
For the study
of the various types of cells the blood
must be dried and stained;

good blood smear for staining
cacy.
and the preparation

is
of a
deli-
a matter of
some
The blood may be obtained from
the
lobe
of

no
the ear

by pricking with a
sterile
lance or glover's needle

skin.
after carefully
washing the surface of the
small
drop should be caught on the edge of a clean
slide,
which
Fig. 43.
lar casts, leucocytes,
Urinary Sediment in Acute Bright's Disease. (Granuand epithelial cells.) (After Ultzmann and
200 diameters.
Hofmann.)
is
then brought in contact with the surface of a second
clean slide.
at
its
The upper glass

is
is
drawn along
the lower one
an acute angle
edges and
so that the blood-drop escapes beneath
spread over the lower slide in a thin
THE MICROSCOPE
even smear.
staining.
IN MEDICINE
AND SANITATION,
is
After drying in the air this
ready for
One
is
of the
most
satisfactory differential blood- stains
Wright's modification of Leishman's method, described

the Journal oj Medical Research for January, 1902
in
(volume VII, p. 138).
The somewhat
complicated pro-
cedure for making this stain and some discussion of

the philosophy of
its
action
must be sought
its
in the original
paper; but the student will find
application easy.
The
dried film
is
covered with the solution of the dye in

is
methyl alcohol for one minute and water
added, drop
by drop,
until
the
mixture becomes
semi-translucent
surface.
and a yellowish metallic scum forms on the

This mixture
for
is
allowed to stand on the preparation

off in distilled
its
two or three minutes and then washed

till
water
the film has a yellowish or pinkish tint in
thinner portions.
The
slides thus stained, dried,
and mounted
in
balsam
should show the various types of blood-cells beautifully

differentiated.
The
red corpuscles, or erythrocytes, are

/x
orange or pink in color and about 7.5
in diameter.
The white
to
cells
may be
divided into three classes accordtheir nuclei

stain.
ing to the relation

the
constituents
cells
which
and granules bear

eosinophils or
of the
The
oxyphiles are
containing matter which takes such
acid stains as eosin.
Stained with the Wright stain, these
appear faintly blue with a dark lilac-colored nucleus
and numerous
the size
large reddish granules.
They
are double
of the erythrocytes.
The
basophiles are cells
which take basic stains
and are
of two varieties.
Small
112

or
basophiles
lymphocytes are about the
size
of
the
red corpuscles, robin's egg blue in color with a round
dark-blue
nucleus and
few
fine
dark-blue granules.
size
Large mononuclear basophiles are three times the

of the red cells
and pale blue with a large oval blue nucleus.
The
the
third type, the neutrophiles, are intermediate between
red-granuled
eosinophiles
are
and
the
blue-granuled
twice
the
basophiles.
size
They
polymorphonuclear,
of
the
erythrocytes,
and
blue, with
one or more
darker lilac-colored nuclei of a twisted shape and numerous
medium-sized
granules
of
reddish-lilac
color.
Blood-plates are almost always seen, purplish rounded

or oval
bodies one-third
the
size
of
the erythrocytes,
with irregular edges and iine blue mottlings.

logic conditions
In patho-
numerous other
cell
elements
may be
used for
for the
present.
Most
of these cell elements are seen in Fig. 44. as that described

is
Such a stained preparation
the study of the structure of the blood-cells
and
determination of the relative

various types.
is
number
of white cells of the

it is
differential
blood count, as
called,
made by counting
several
hundred white
cells
and
calculating the percentage of each form.
For thus de-
termining the absolute number of the blood corpuscles the
apparatus called a hsemocytometer

sists of
is
designed.
It
con-
a glass slide containing a cell of

of o.i
known diameter
is
and a depth
mm., on
the bottom of which

off
ruled
a micrometer scale marking

side.
squares 0.025 rnm- on a
The blood
the
cc.
is
diluted, in a special pipette constructed
for
purpose,
with
100 parts of Gower's

g.
solution
(100
water, 15 cc. acetic acid, 6
sodium sulphate),

THE MICROSCOPE IN MEDICINE AND SANITATION. 113
when red
cells the
cells
arc to be counted.
is
For counting white
blood
diluted with 10 parts of
\%
acetic acid
cells
tinted with gentian violet,
which destroys the red
and
faintly stains the white corpuscles.
Wide
variations of the blood elements, both in absolute

in proportions, occur in
numbers and
many normal and
Fig. 44.
Blood-cells (Wright's Stains). (Photomicrograph by W. C. Greene, Harvard Medical Schoo|.) (Erythrocytes, large basophiles, polymorphonuclear neutrophiles, and blood plates.) 700 diameters.
abnormal conditions.
The blood
of the adult
human being
and 3000
to
should contain 5,000,000 erythrocytes per cubic millimeter

in the
male and 4,500,000
in the female
10,000 white blood-cells, of
which 70-72%
should be
polymorphonuclear neutrophiles, 25-30% basophiles, and
1%
eosinophiles.
In anaemia the red
cells
may
fall to
360,000 per cubic millimeter and in other conditions they
may much
exceed the normal.
Leucocytosis, or the in-
114
ELEMENTS OF APPLIED MICSOSCOPY.

cells,
crease of white
accompanies
all
acute inflamma-
tions as well as certain other conditions
and may
affect
any one type of

it
cells
or
all of
them.
In inflammations
is
the
polymorphonuclear
neutrophiles
which
are
increased; in trichinosis, on the other hand, the eosinophiles

5.
may
reach
50%
of the total.
The Detection
are
of the Malarial Parasite.
Blood
ex-
aminations
malaria.
to the
also
undertaken for the
diagnosis
of
The Protozoon
malarial parasites gain entrance
body from the
bite of
an infected mosquito
in the
form of minute spore-like bodies which penetrate the

red corpuscles and there assume a crescentic form.
Later
the organism becomes amoeboid, sends out blunt pseudo-
podia, and develops nuclear granules.
As
it
matures,
it
occupies the greater part of the red corpuscle and finally

begins to divide, appearing in a rosette form, at last separating
and discharging
into the blood stream granules
like those
with which the cycle began.
Blood
may
be examined in
its
fresh condition for the
malarial parasite, but better preparations are
obtained
yields
by
staining.
The Wright-Leishman method

the
ad-
mirable
results,
cytoplasm of the parasite

color varying
being
stained blue
lilac
and the nuclear granules a
from
through red to almost black.

of blood-cells
The
general appear-
ance
containing the parasite of tertian

is
malaria,
HcBmamceba vivax,
45
;
indicated in the two upper
fields of Fig.
the lower field shows a later flagellated
stage in the flood stream.

in the examination of
Only considerable experience

will enable the ob-
normal blood
server to recognize the invading organism with certainty.

THE MICROSCOPE
IN MEDICINE
AND
SANITATION. 115
The
presence in the blood of free "pigment," probably
stainable material
sites, is
from decomposed blood- cells or para-
characteristic of malaria.
Fig. 45.
^The Malarial Parasite (Stained).

Spitta.)
(After
Slater
and
1000 diameters.
6.
The Examination
of
Sputum
for Tubercle Bacilli,
and the Cultural Diagnosis
of Diphtheria.
Besides
the
Ii6
examination of blood smears for malaria the bacteriologist of
a Board of Health
is
expected to
make
diagnoses
of diphtheria
and
tuberculosis,
which are of
special' im-
portance on account of the necessity for prompt treat-
ment
of the former disease
and on account
of the difficulty
latter.
sometimes experienced in recognizing the
Both
maladies are due to the presence of bacteria, very minute
rod-shaped fungi which
first
gain lodgment in the respira-
tory or alimentary passages.
In the case of consumption or pulmonary tuberculosis

the bacteria are discharged in
enormous numbers
in the
sputum and may
there be detected without great difficulty.
When
teria,
stained with an anilin-dye the tubercle bacilli are
not decolorized by dilute sulphuric acid as are most bac-
and
this property is
made
the basis for a simple

thick smear of
method
is is
of differential staining.
sputum
This
made on a
glass slide
and dried over the flame.

it
stained for two minutes, heating until

(i
steams, with
Ziehl-Neelsen's carbol fuchsin
gram
basic fuchsin in
ID
cc.
95%
alcohol mixed with 90 cc. of a
5%
aqueous
solution of phenol).
The
slide is
then washed under the
tap and immersed for two minutes in a solution of 3 parts

of hydrochloric acid in 100 parts of
95%
alcohol.
It is
then washed again and counterstained- for half a minute

in a
dried,
1%
aqueous solution of methylene blue, washed,
and mounted.
The
slender tubercle bacilli should
be stained bright red with fuchsin, while other bacteria
and the leucocytes and other

colored faintly blue.
cell
elements present are
In
all
work with
bacteria, the V^-inch objective
must
THE MICROSCOPE IN MEDICINE y4ND SANITATION,
n?
be used with a drop of cedar- oil between the lens and

the cover-glass, or the slide,
if
no cover-glass be used.
The Abbe condenser

terial preparations.
is
also a necessity for lighting bac-
The
diagnosis of diphtheria
is
somewhat more comand involves

cultural
plicated than that of tuberculosis

as well as microscopical methods.
at the
is
The organism
occurs
back of the throat and material

sterile
for examination
obtained by rubbing a
swab
of cotton over the
whitish patches which appear on the pharynx and fauces.
This swab
is
then passed over a slanting surface of coag-
ulated blood- serum

to twelve
upon which
will
after incubation for six
hours at the body temperature minute pearly

of the
bacilli
colonies
appear.
sterile
portion of the
growth
is
then removed with a
loop of platinum
wire and spread over a clean cover-slip in a drop of

water.
The
cover thus prepared
is
dried, fixed
by pass-
ing three times through the flame, and stained for ten
minutes
with
Loeffler's
methylene blue
(30
cc.
of
a
cc.
saturated alcoholic solution of the dye mixed with 100

of a Vio.ooo
aqueous solution of caustic potash).
After
washing,
drying,
and mounting
in
balsam, the specivariable
men,
in cases of diphtheria, will of
show the very
rods characteristic
the diphtheria bacillus,
many
of
them cigar-shaped or club-shaped, and

Sometimes
these
often exhibiting
the peculiar cross-barred appearance indicated in Fig.

46.
organisms
will
be
seen
mixed
with spherical bacteria arranged in pairs and chains,

the
streptococci.
In negative
cultures
only cocci are
found,
as a rule, although
other baciUi,
more or
less

ii8
closely

resembling
the
diphtheria
organism,
may be
present.
7.
The Serum Reaction

most
important
progress
in
Typhoid Fever.
results
One
of
the
practical in
of
the
rscent
marvellous
of
the
study of
the
phenomena
immunity has been a method

of
for the
clinical diagIt
nosis
typhoid fever and certain other diseases.
Fig. 46.
Diphtheria Bacilli
brook.)
(Methylene Blue).
Diagrammatic.
(After
Wes-
has been shown that when the animal body
is
invaded
by a
cases
parasitic micro-organism, the cells
produce in some
antitoxins
which neutralize the poisons of the
microbe, and in other cases anti-bodies of another type
which have a
itself.
specific destructive action
on the parasite
Sometimes the defensive secretions dissolve the
THE MICROSCOPE
foreign cells;
IN MEDICINE
AND SANITATION.
119
sometimes they cause them to clump
to-
gether in masses and to settle out of the fluid in which
they are suspended.
In the blood of an individual
in-
fected with typhoid fever, bodies of the last type,
known
is
as agglutinins, are present
and
their
clumping reaction
It
is
specific for that particular micro-organism.
true that
the blood of a normal individual
may
contain substances
which agglutinate typhoid

patient has the
bacilli,
but that of a typhoid

it
power
to
such a high degree that
will
produce the same

dilution.
effect
much more
is
rapidly
and
in high
In making the diagnosis of a suspected typhoid

taken from the lobe of the ear
to clot in a small test-
case,
a sample of blood
in the
usual manner and allowed
tube so that the corpuscles are separated from the clear

straw-colored serum.
drop of serum
is
then mixed
on a
slide
with 40 drops of a fresh broth culture of the
typhoid bacillus.
The mixture
is
covered and examined

objective.
under the i;V-inch oil-immersion

bacilli
At
first
the
may be
seen, after careful focussing, as
minute trans-
parent rods, singly, or in pairs and short chains, moving

rapidly about across the field of the microscope.
tinins
If agglu-
be present, the bacteria become motionless and clump
together in masses, sometimes large enough to be visible

to the
naked
eye.
If the reaction is not obtained
with
a 5V dilution, another drop of serum

if this fails,
may
be added, and
should
again, another.
bacilli in fifteen
The
latter dilution, iVi

if
clump the
be needed.
minutes
the case examined
be typhoid fever; with a 4V dilution an hour's time
may
The
test
may be made
quite as well with dried
blood after diluting with the proper amount of water.
I20
8.
ELEMENTS OF yiPPUED MICROSCOPY.

Tho Microscopic Examination
trichinosis
of
is.
Pork
for Trichina.
The disease known as

typhoid fever caused
Trichina
is
hke diphtheria and

micro-organism,
by a
parasitic
although the parasite belongs to a very different group.

a minute worm, barely visible as
a. speck
to the
naked
eye,
which bores
itself in
its
way
into the muscles of swine
and there encysts
a calcareous nodule. If such pork

is
be eaten, imperfectly cooked, the limy cyst

the
dissolved;
worms emerge and
reproduce, and a myriad of their

tissues of the body, causing If the patient recovers
progeny penetrate the
high
fever and, sometimes death.

this crisis, the
from
worms
encyst themselves in the muscles

serious difficulty.
is
and produce no further
In America, trichinosis
rare in
it,
man, although some

is
2%
of swine are affected with
because pork
eating.
more
or less thoroughly cooked before
In Europe,
however, where pork products are eaten almost raw, the
danger
is
serious
and must be met by preventing the

Elaborate governinstituted in
distribution of trichinous swine flesh.
ment systems
of
meat inspection have been

in Prussia,
for example,
many
States
countries;
over 25,000
officials
are employed for this purpose. the
In the United
for
Department
of Agriculture
some years
com-
maintained a bureau of meat inspection which examined

all
pork intended for foreign export and
interstate
merce, employing a large corps of microscopists.
Samples of pork
to
be examined for Trichina are taken

in pieces
from the diaphragm or other muscles and cut

about an inch by half an inch in
is
size.
One such
piece
then placed between two slides of heavy glass, mounted

in a
121
frame and provided with screws, by which they can
be pressed together, thus reducing the meat to a thin

transparent layer.
The frame
fit
bears on
its
under side
two ridges which
into grooves in a special corrugated
stage of large size on the microscopes designed for this
purpose.
Under a very low
objective, of ij-inch equivais
lent focus, the

slid
compressed frame
its
then examined, being
along until
whole width has been covered, and then
shifted to the next groove, so that a
new
field
may be
examined.
In
this
way
the whole sample

observer.
is
may
be viewed
in five minutes
by a trained
Under
the microscope the pork
seen to be
made up
of the long cylindrical cross-striated fibres of voluntary
muscle, and
if
Trichina be present the cysts will be seen
here and there as spindle-shaped, whitish bodies.

sections of trichinous
In
pork prepared
after paraffin im-
bedding with the microtome, the worms may be seen

coiled
9.
up inside the cysts, as shown in Fig. 47. The Microscopy of Drinking "Water. The
sanitary
quaHty of water depends primarily upon the presence

or
absence of disease
germs,
generally
introduced in
its
sewage.
is
The most important
evidence as to
character
therefore obtained
by the bacteriological and chemical
analyses,
which indicate with great delicacy the presence
of minute traces of sewage pollution.
microscopical
of
examination may,
value by
cells,
however,
often
add information
showing the presence of starch-grains, yeasttissue,
fragments of vegetable
and
certain Infusoria
characteristic of
decomposing material; organisms pecul-
iar to a particular
pond or stream have sometimes proved

122

its
of importance in the detection of admixture of
water
with that from some other source.
The
great importance
lies,
of the microscopical examination of drinking-water
however, in
its
application to the study of those organisms

tastes
which produce
tastes
and odors
in
reservoirs.
Such
and odors are
in almost all cases
due to the mul-
FiG. 47.
Trichina.
(After Hager-Mez.)
50 diameters.
tiplication of microscopic plants
and animals belonging
to the Algje
and the Protozoa; and the study and control

depends upon the systematic use of
of these organisms
the microscope.
considerable concentration of the sample must, of
course, precede the microscopical examination of drink-
ing-water;
and
this is usually
fine*
accomplished by
filtration
through a layer of
sand.
The Sedgwick-R after

the
method, in routine use by the State Board of Health and

the
Metropolitan Water Board of Massachusetts,
THE MICROSCOPE IN MEDICINE /1ND SANITATION.

Water Department
of
123
Brooklyn, and
other
sanitary
authorities, is as follows:
measured quantity of water,

through perhaps a quarter
circle of bolting-cloth suptall
half a liter or a
of
liter, is filtered
an inch
of fine
sand on a
ported by a perforated rubber stopper in a
funnel.
The sand
with the organisms collected on
it is
dropped
of water,
is
into a test-tube, shaken
up with a small volume

a moment.
of
and allowed
poured
to
off
to settle for
The water
then
and a second portion
wash-water serves
remove almost aU the organisms from the sand.

to
The
some
two washings mixed together are then made up
convenient volume, 10 or 15 cc, and thus one has in say
10
cc.
the organisms originally present in a
liter.
One
cubic centimeter of the concentrate

i
is
then placed
in a cell,
mm.
deep and 1000

is
sq.
mm.
in area.
special
type of ocular micrometer
used with a square ruled
upon
it,
and the lenses and draw- tube are so adjusted

i
that the square shall coincide with
sq.
mm. on
the stage.
cell
Ten
then
or twenty fields in different parts of the
are
examined and the organisms counted,
when a
number
simple calculation enables one to compute the

present in
i cc.
of the original sample.

of micro-organ-
By
this
method the kind and number
isms present
may be
The
easily
determined, the cause of

of future
diffi-
existing trouble detected,

culties predicted.
and the prospect
efhcacy of methods of
purifij:a-
tion may be tested and the source of trouble located the particular part of a supply where it exists.
in
few of the most serious odor-producing organisms
are figured in Fig. 48.
At 4
is
Anabcena, the Blue-green

124

city of Springfield,
Alga which has cost the
Mass.,
many
a
thousands of dollars by the production in Ludlow Reservoir of
the
"pig-pen" odor.
At
is
Asterionella,
Diatom which caused
the geranium odor in the water of
Fig. 48.
Micro-organisms of Drinking-water.
Whipple.)
1, 2, 3.
(Redrawn from
4, 5.
300 diameters. Asterionella. 6. Synura. Anabaena. 7. Uroglena.
Brooklyn, N. Y.
The
Protozoon, Synura, at 6 was the
author of the cucumber taste in the Boston supply,
and
of
its ally,
Uroglena, at
7, is
perhaps the worst offender
all,
having caused trouble in some 44 supplies in Massa-
chusetts alpn^,
The microscope in Medicine /ind sanitation. t^5
REFERENCES.
Cabot, R. C.
1903.
Clinical
Examination of the Blood.

of Disease.
New New
York, York,
Cabot, R. C.
1899.
The Serum Diagnosis

Protozoa and Disease.
Clark, C. EwiNG, J.
J.
New
York, 1903.
Phila.,
Clinical Pathology of the Blood.

J.
1903.
McFarland,
MuiE,
R.,
Text-book upon the Pathogenic Bacteria.
Phila., 1900.
and Ritchie, J. Manual of Bacteriology. Amer. by Harris, N. M. New York, 1903. Oertel, T. E. Medical Microscopy. Phila., 1902. PuRDY, C. W. Practical Uranalysis and Urinary Diagnosis.
ed.,
Phila., 1895.
J.
Tyson,
Guide
to the Practical
Examination of Urine.
Phila., 1902.
Whipple, G. C.
York, 1899.
The Microscopy
of Drinking-water.
New
Wright,
J.
H.
of Blood Films
Rapid Method for the Differential Staining and Malarial Parasites. Journal of Medical
Research, VII, 1902, 138.
CHAPTER
X.
FORENSIC MICROSCOPY.
1.
The Microscope
liê
in
Law.
In
in
a court of law where
questions of
and death may hang upon the nature

study the minuter
identification
of a blood-stain or the genuineness of a document, the
microscopist
structures
of
is
often
called
to
disputed
objects.
The
of
persons by the anthropometric system of Bertillon
may
be supplemented to advantage by a detailed examination

of the ridges
upon
the fingers.
The
detection of
human
be
blood
is
greatly facilitated
by microscopical methods.
In sexual cases the presence of spermatozoa

easily
may
determined under the microscope.
Low
powers
aid in
the study of bullet-wounds with respect to the
distance
the
as
and
direction
etc.
from which the shot was
fired,
powder used,
regards
the
The examination
writing
of documents,
paper and
utensil
used,
the
presence of erasures or alterations, and the

istics
characterfield
of
the
handwriting,
is
an important
for
microscopy.
2.
The Examination
is
of Blood Stains.
Frequently the
minute
J26
expert
stains
asked to decide as to the character of bloodclothing, furniture, etc., stains often of
upon
size
127
and sometimes
of considerable age.
Two
as to
problems
are involved
first,
as to the presence or absence of blood
in the specimen presented,
and second,
its
nature,
whether
general
human may be
or animal.
The
presence of blood in
settled pretty easily
by four methods
one
one
chemical,
one
microchemical,
microspectro-
scopical,
and one purely microscopical.
The chemical
an alcoholic
test consists in the application to the stain of
solution
of
gum
guaiacum, which yields a blue color

of aerated turpentine.
fact that haemo-
reaction
on the further fddition

test
The microchemical
globin, the
depends on the
complex albuminoid substance characteristic
of the
red blood-corpuscles, yields on treatment with
acetic acid a crystalline iron salt, haematin,
whose
triple
chloride
is
known
as hsemin.
is
In making
this test the
blood,
if
in a dry condition,
placed on a slide and gently
warmed
rate
in glacial acetic acid.
This
is
allowed to evapo-
and replaced by a 0.07%

in its turn
is
solution of
sodium
chloride,
which
carefully evaporated without heating
the slide above 60 C.
Another drop of
acetic acid
is
added and the

boils.
slide is this
time heated so that the liquid

is
After evaporation
if
it
examined under the
^-in.
objective;
blood be present, crystals of haemin (or

flat
haematin trichloride) will appear as minute
rhom-
boids often superposed in the form of a cross.
They
vary from yellow to reddish brown in color and from

0.005-0.02
mm.
in size.
This
test is
a delicate one, serv-
ing to detect from 0.05-0.15 mg. of dried blood, and even
very old material gives satisfactory results, as in the classic

case of the blood of the murdered Kotzebue, detected
128

on the papers which lay upon
sixty years after his death

his desk.
For
examination
by
the
microspectroscope,
distilled
fresh
blood-stains are treated with
water and older
specimens with weak acetic or

in a small glass cell
citric acid.
"When placed
to
and examined by the methods
be
described in the next' section, the characteristic spectrum

of haemoglobin or one of
its
derivatives will be apparent.
The
and
direct microscopical examination of blood-stains

at
aims simply
serves
the
detection
of
the
red
corpuscles
that
to distinguish
mammalian blood from
of birds, the for the
three
tests
above outlined being general
hemoglobin
blood
of all red-blooded animals.
Dried
and and
clotted
must
first
be
treated
with
some
solvent which shall attack

set
the
albuminous coagulum
free
the blood-cells.
Numerous media have

solution of potassium
been suggested, of which a

hydrate
is
33%
one of the simplest, and Ranvier's solution

2
(potassium iodide,
of iodin, 100 parts)
parts;
is
saturated aqueous solution
one of the best.
In any case a
bit of clot or the scraping
from a
stain is covered with
the solvent
scope.
and placed
in a hollow slide
under the micro-
Sometimes
after a
few minutes, sometimes only

clearer
after days, the opaque
mass becomes
and the
individual cells appear as circular, non-nucleated discs

7.5/1
in
diameter.
Under most conditions the
discs
appear somewhat biconcave, with a thickened ring about

the edge.
In the original process of drying or in the treatment

with a solvent, the blood-cells
may
be so distorted that
their
129
recognition
is
not easy.
In water, for example,
the corpuscles swell
up
to a spherical form.
The comlitera-
monest objects which
may be mistaken
instances
of
for blood-cells
are the spores of certain plants,

ture
and medico-legal
erroneous
contains
ludicrous
expert
opinions based on such findings.

tion carefully
With a good preparaof fresh blood,
compared with specimens
such errors can easily be avoided;
and, since the cor-
puscles of birds are of oval shape and

nucleus, blood-stains of
show a
distinct
avian origin
may be
excluded.
The blood
that of
of other
mammals can be
differentiated
from
man
only by the size of the
cells,
and the
differ-
ences are so slight that a certain result can seldom be

reached.
The blood
of the goat has cells less than 4.5
//
in diameter, while the corpuscles of the horse
and cow
and pig are under 6

rabbit, closely
fi;
but the corpuscles of the dog,
mouse,
cat,
and other domestic animals very
resemble those of
human
blood.
Such
differ-
ences of one or two micromillimeters cannot be relied
upon, since even in the fresh blood of a living animal
some variation occurs and

are
distorted
dried
specimens
much more
dubious.
Dr. E. L. Walker has pointed
out that the white blood-cells of different
mammals
differ
much more
widely than do the red corpuscles;
and the
study of these bodies

microscopist.
may
furnish valuable aid to the
The
crucial test for blood-stains,
and the only method
by which
the blood of
man may be
definitely distinguished
specific
from that of other mammals, depends upon a

biologic reaction
similar to that
which
is
used in the
13

test for
serum
typhoid fever.
If
human
blood- serum be
injected into the
lymph spaces
of a rabbit, there appears
in the blood of the rabbit a substance
which
precipitates
this reac-
certain albuminoids in the blood of

tion serves to distinguish
man; and
human blood from
that of the
cor-
domestic animals with certainty.

puscles of apes
Only with the

of the
test,
and monkeys does the serum
immuwhich
nized rabbit react in a similar manner.

ordinarily bears the
it,
This
first
name
of Bordet,
who
suggested
is
not applied under the microscope, but in small tubes

is
in
which the precipitation

3.
detected macroscopically.
Its
The Microspectroscope and

and
in
Use.
În
testing
blood-stains,
many
other fields of microbiology

aid.
and microchemistry, the spectroscope may furnish
As
as
ordinarily used,
the instrument consists of a tube

of a microscope
fitting
on the eyepiece
and containing,
flint
shown
in Fig. 49, a triangular prism of

light
glass
which disperses white
into
its
constituent colors
and two prisms

tion,
of
crown
glass set in the opposite direc-
which serve
first
to counteract the refraction
produced
by
the
prism.
The
principle
is
the
same as that
the latter
of the achromatic objective, but whereas in
the angles are arranged to produce a net refraction with
neutralized
dispersion,
the
prisms of the spectroscope
yield a net dispersion with practically
no
refraction.
The
special
type
of
eyepiece used with the micro-
spectroscope contains a diaphragm which only allows a

longitudinal
slit is slit
of light to pass,
and the width
of the
for use
regulated by a set screw, being cut
down
to half a millimeter or less.
When
the detachable prism

tube
is
I3f
well lighted, a
placed in position and the
field
its
spectrum should be seen lying with

right angles to the
slit
colored bands at
If the
in the ocular.
spectrum
lines are oblique, the

it
prism tube must be rotated until

slit.
bears the proper relation to the
Reference to Fig. 49 will show also a lateral tube connected with the spectroscope, into which rays of light
Fig. 49.
The
Microspectroscope.
(Bausch and Lomb.)
from outside are projected by a mirror, and a prism
by which these rays are
reflected
upward.
This
is
used
when
light
it
is
desirable to
compare the spectrum
of sunof the
with that of some object upon the stage
microscope.
When
properly adjusted the two spectra,

light
produced respectively by
microscope and by
will
passing up through the
that reflected through the side tube,
appear
to lie side
by
side.
132
ELBMENTS OF APPLIED MICROSCOPY.

sensation of light
is
The
tions,
produced by the ether vibra271

T-KK
whose length lies between
and
10,000,000
10,000,000
of an inch.
it
When
any
solid
body
is
gradually heated
becomes luminous,
first
emitting the longer red rays

later
at
a rate of 458 million of millions per second and
the rays of other colors as well, culminating with the violet
rays,
whose
rate
is
727 million of millions.
At
this
point the
all colors
body has reached a white

are given
off.
heat, since rays of
If
now
the white light from such
an incandescent body be passed through the spectroscope,

it
is
broken up, through the different refrangibihty
of
the rays of different amplitude, into a continuous spectrum

in
which
all
the rainbow colors appear merging into each
ether.
An
incandescent gas, on the other hand, profield
duces a line spectrum, most of the
being dark, with
here and there narrow bright lines whose
number and
position are characteristic of the particular substance.
The
color of objects
is
due
to their property of transIf
mitting or reflecting rays of a certain amplitude.
white light be passed through certain

gases
solids, liquids, or
below
their
point
of
incandescence,
and then
through the spectroscope, the presence of black bands

crossing the spectrum shows that light of certain definite
wave-lengths has been removed.
gas cuts out the
same light-waves which

temperature.
it
itself
produces at a higher
The spectrum produced by

one, like
that
sunlight
is
a continuous
solids,
produced by other incandescent
crossed here and there by dark bands, the Frauenhofer
lines,
133
which are due
to the presence of the
cooHng vapors
of metallic elements in the surrounding atmosphere, each of
which absorbs
the
light
corresponding to
its
own
con-
bright line spectrum.
The Frauenhofer lines may be
sidered the shadows of the spectra of the metals in the
outer atmosphere of the sun.
Obviously
artificial light
should be used for the micro-
spectroscope, since the Frauenhofer lines would intro-
duce
confusing
element.
When
various
solid
and
liquid substances are placed
on the stage
of the micro-
scope, absorption spectra of the
same general type
as
that of sunlight are produced, although the dark bands
are wider
and
less
sharply defined, this being more and

of the
more the case as the thickness

is
body
is
increased.
It
therefore desirable to examine layers of varying depth,

this
and
may
slide
it
be done by means of a
cell
made by cementoff obliquely
ing to a
so that
a piece of heavy glass tubing cut
shall
be
mm.
deep on one side and 0.5
mm.
deep on the other.
The
ination
application of the microspectroscope to the examof
various
organic
bodies
its
has
not
yet
It
been
has
developed to anything Hke
possible limits.
yielded interesting results in the study of certain anima L
and vegetable coloring-matters;

coloring-matter
of
but
its
chief
use has
so far been in the detection of blood-stains, since the
red
blood
produces
characteristic
spectra..
Suspected blood-stains which are to be thus

treated with distilled water until a
is
examined may be
reddish-brown solution
that
produced,
when
it
is
apparent
liquid
haemoglobin has gone into solution.
The

134
ELEMENTS OF
/APPLIED MICROSCOPY.
in
thus obtained,
when placed
with
cell,
covered and exof
amined with the low

oxyhsemoglobin
green,
objective,
shows the spectrum

in
dark bands
the
yellow
and
shown
in Fig. 50 at
4.
If
a drop of
ammonium
sulphide be added, the two bands will fuse into one, giving
Fig. 50.
Spectra of Oxyhsemoglobin and reduced ILemoglobin.

(After Howell.)
the
spectrum of
addition of a
reduced
little
haemoglobin,
shown
at
B.
The
citric
acid to a solution of oxyacid haematin,
hsemoglobin produces the spectrum of

with a broad band
in the red, a
narrow one in the green,

In old blood-stains
to hsematin,
and a very
faint
band
in the blue.
the oxyhsemoglobin has been changed

since
this
and
substance
is
insoluble
in water,
is
no
color-
ation of the liquid appears
when
the stain
treated as
tried;
above.
In such a case a
little
acetic acid
must be
at
this dissolves hasmatin,
and the solution
once pro-
duces the acid haematin spectrum.
On
adding
am-
monia, the broad band in the red disappears.

stains in
Blood-
which the hsemoglobin
is
only partially changed
to
13S
hasmatin
(methsemoglobin)
with the combined bands of both.

test
is,
may show The

of
spectrum
spectroscopic
of course, given
It is
by the blood
any red-blooded
animal.
extremely delicate and has yielded posiold.
tive results
4.
with stains over a hundred years

of Finger-prints.
The Study
The
cases,
identification of
individuals, so important not only in criminology but in
a multitude of
facilitated
tillon.
civil
and criminal
has been greatly

of
by the anthropometric system

rests
M.
Ber-
This
upon the
principle that variations in
the proportions of the body,
when a number
of separate
features are measured, are so characteristic that a definite
formula
may
be obtained from them, which shall
differentiate
each individual from any other.
Commonly
foot
the length and breadth of the head and the length of the
left
middle
finger, the left forearm,
and the
left
form
a primary basis for

characteristics
classification to
which other general
and
special
individual peculiarities
may
be added ad infinitum.
The
to
use of the finer details of the structure of the skin

first
supplement the Bertillon system was
suggested
by the distinguished English

though
Sir
biologist, Francis
Galton,
finger-
William Herschel had previously used
prints as a
means
of personal identification.
Recently
Professor H. H. Wilder's investigations have shown the

great importance of
heredity.
If
these
structures in
the
study of
the bulb below
the tip of the finger be

thin
pressed
first
upon a metal surface covered with a

record will
film of printer's ink,
and then upon a paper or card,

be obtained of the arrangement
a permanent
136

and furrows
characteristic of that digit, a
of the ridges
record necessarily free from the error which
ordinary
anthropometric
measurements.
may attend GaUon has

unchanged
shown
that the pattern of the skin persists

life
through
and
is
unaltered in
its
essentials
by
cuts,
burns, or any ordinary accidents.
The
variations are so,
great that he calculated the chances of identity between
two
single
prints to be only one in sixty-four billion.

is
When
ten digits are compared, identification

records, in order to be of
absolute.
service,
Such
any practical
this
must be readily arranged and
classified;
Galton
2
(Redrawn
'
after Galton.)
Fig. 51.
F1NGEE.-PKINT Patterns.
has done so successfully that, out of a "finger-print directory" of 2632 persons, any pattern could be located in
three minutes.
In the
first
place, the patterns of the skin
may
be grouped under three heads.
In
all
cases the
papillary ridges run across the fingers in the vicinity

of the third joint,
and
at the tip they follow the curve
of the nail in a rounded arch.
Sometimes the ridges

less
between follow the outer ones in a more or

arch of lessening convexity; this
is
even
the arch type
(i.
Fig. 51).
Sometimes the intermediate ridges form a loop running
from one
side
inward
to the center of the
bulb and then
doubling back again.
Obviously at the opposite side
of the finger the outer ridges, the loop,
137
and the basal
ridges will cut off between
them a
is
triangle,
known
as
the delta
(2,
Fig.
51);
this
the loop type.
Finally,
the ridges on the bulb
may be
two
so twisted as to form a
complete
circle cutting off
deltas,
one on either side;
this is the
whorl type
(3, Fig. 51).
classification
of the
ten digits according to their

serves for a primary division
arches, loops,
and whorls
of finger-print cards into a
number
of general classes.
Since, however, certain combinations of these digital for-
mulae are
division
much more common

;
than others, further sub-
must be made
and
this
may
be accomplished by
studying the minuter structures of the finger-print, and

particularly
digits
by counting the
ridges in the loops of those

It
is
which show that
structure.
here that the
microscope comes into play, since such details cannot

well be
made
out with the naked eye.

is,
magnification
of only ten or twenty diameters

sirable;
however, generally de-
hence the instrument used must be either a simple
microscope, preferably mounted on a stand, or a com-
pound microscope of very low power and very wide field. În the study of 5. The Examination of Documents.
disputed documents the microscope

ble service to the legal expert.
may
and
be of considera-
The
material of which
its
paper
is
made,
its
texture
and
sizing,
water-marks
instru-
should
first
be noted, the character of the writing

it
ment and the grooves
cuts
upon the paper,

Inks
if
steel
pen, being possibly significant.
various chemical methods; but
may be tested by much may be learned
under the microscope from an observation of the color
138

luster of the deposit
and metallic
taining
formed by inks con-
gums and
of the solid particles of coloring-matter
always present.
Even
the changes of consistency in the

this
same
bottle
on standing may be detected by
method.
Direct evidence of forgery in the shape of alterations,

erasures, or interlineations
is
often furnished
by exam-
ination with a
hand
lens
or
low-power compound
sizing
microscope.
Erasures remove the
and loading
material of the paper and leave loose ends of teased-out

fibres
in
which the ink of
later
writings runs freely.
Marks
in
of preliminary tracings are sometimes apparent of

it
cases
elaborate
forgery.
When
two
lines
cross
each other
may
be of importance to determine which
was made
service,
first;
and
here,
line
too,
the microscope
is
of
since the
upper
often shows a widening,

line,
due
to capillarity,
on entering the lower

it.
and a nar-
rowing on leaving
The
continuity of the pen fur-
rows of the upper
line
may
also
be apparent.
Obliquely
viewing the point of intersection with a hand lens, along

the two lines successively, helps to
make
itself,
it
clear
which
one was superposed.

Proceeding
spacing of the
to
lines,
the
handwriting
the
first
general
words, and letters should

the shading,
be noted.
The pen
of words
pressure,
the general
symmetry
and letters, and the firmness
of individual lines are

itself in
all significant.
Temperament manifests
haste or
caution, in energy or reserve.

feebleness,
Lack
of facility, physical
and the labored attempt
to imitate, alike pro-
duce
tremors which
may
easUy be distinguished from
each other.
FORENSIC MICROSCOPY
All these obvious peculiarities
skilful
139
imitated
may be
by a
hand, and
it
is
only more minute characteristics

identification of handwriting.
which serve for the certain
-2
Fig. 52.
^MnsruTE Characteristics of Handwriting Lines.

(After Frazer.)
The
latter are
studied by two methods, the statistical

First, the
and the microscopical.
measurements of the
distance between certain letters in a given word, of the

ratio of heights to breadths in certain letters,
and
of the
Angular glope of certain long strpkeS; when averaged.
14

from which the deviations
of the writer are
etc.,
yield values
rarely great.
Composite photographs of signatures,
are important aids in

place,
work
of this sort.
In the second
more minute
differences exist
which can be detected
only under the
compound microscope.
any long
lines
These are of three
magnitudes.
First,
show
certain varia-
tions of direction
whose number and nature vary with
the writer.
Second,
much
from
finer fluctuations occur, visible
only under the

vertical
compound microscope,
side
to
in the shape of
deviations
side
and changes
in
width due to periodic changes in the pressure of the penpoint.

still
Third, on one or both margins of the line are

lateral serrations.
more minute
These
vertical
52.
and
lateral variations are

are,
both illustrated in Fig.
They
of course,
influenced
more or
less
by paper and
and number
writing utensils
and by the physical and mental condiyet the size, position,

of
tion of the writer;
the waves, swellings,
and notches are so related
to the
nervous organization of the writer, and differ so markedly

with different individuals, as to yield important aid in
the identification of handwriting.
REFERENCES.
Babcock,
J. F.
other Stains, Hairs, and Fibres.
Frazer, p.
of Blood and York, 1894. Bibliotecs, or the Study of Documents. Phila-
The Medico-Legal Examination
New
delphia, 1901.
Galton, F. Finger Prints. London, 1892. Galton, F. Finger-print Directories. London, 1895. RoscoE, H. E. Spectrum Analysis. London, 1870. Taylor, A. S. A Manual of Medical Jurisprudence. New York and Philadelphia, 1897. Wood, E. S., Withatjs and Becker. Medical Jurisprudence, Forensic Medicine and Toxicology. New York, 1894.
CHAPTER
XI.
MICROCHEMISTRY.
I.
The Application
fields
of Microchemical Analysis.
In
few
have the
possibilities of the
microscope been
less fully realized
than in pure and applied chemistry.
science which deals with the character
and behavior
of chemical substances should surely take into account
the characteristic crystals of which, under certain conditions,
many
such
to
substances
are
composed.
reactions
Yet and
chemists
trained
study gross
color
precipitates have too often regarded the microscope as
strange
and impracticable instrument. and elaborate secondary

been neglected.
tests
Systems
of
recondite
have been
built
up, while the direct study of fundamental physical characters has
Although the toxicologist and

microchemical
fields
the medical microscopist have used
tests
with such good results that in those special

are
of
they
these
importance,
in
is
general
slow;
the
adoption of
methods by chemists
and only the develop-
ment
of
petrography,
the
microscopic study of rock
sections,
has
at last,
during very recent years, called
general attention to their importance.
The
study of microchemistry
is
by no means
limited
to the examination of crystalline forms alone, as might

141
142

its
be supposed from a perusal of

published by Harting in 1866.
others have since
earliest
exposition,
Lehmann, Behrens, and
shown
that
many dynamic
properties,
melting and volatilization for example, as well as the

typical
chemical reactions,
may
often be observed to
great
advantage
The
use
of
minute quantities of material (one-tenth of a milligram

often sufficing), the rapidity with which residts
may be
obtained,
and the simplicity and compactness of the ap-
paratus needed are general advantages of the microchemical
method, which apply in
all
maimer
of analyses.
On
the whole, few chemists
who
take the trouble to familiarize

fail to find
themselves with the use of the microscope wiU

it
at times
a valuable aid in their work.
In the study
it
of certain closely related organic
compounds
furnishes
at once information which can otherwise be attained,

if
at
2.
all,
only by the expense of a vast amount of labor.

of Typical Crystals.
The Study
student
of
Ît
will
be well for
himself
the
first
microchemistry
to
familiarize
with the most important characteristics used in the
identification of crystals,
by the examination
of certain
typical forms.
The
actual process of deposition should
be studied and those differences observed which occur in

the formation of
as,
crystals
under
different
conditions,
solutions,
for instance,
from alcoholic and aqueous
and by slow spontaneous drying on the one hand and

rapid evaporation by heat on the other.
in a mother-liquor of the
Crystals formed
their
same composition show

and the
characteristics with special distinctness.
The
linear dimensions
relative proportions of
MICROCHEMISTRY.
I43
the various faces of a crystal vary widely with the supply

of material in the solution
from which they were deposited.
The
ever,
interfacial angles
between corresponding faces and
the facial angles between corresponding e'dges are, how-
always constant for a given substance.

as the
This law,
known
Law of
Steno, forms the basis for determina-
tive crystallography,
and makes
it
possible to distinguish
the crystals of such
compounds
as are not isomorphous.
Besides the general crystalline form, there are numerous

special points to be noted in the study of a crystal.
Its
habitus (the size and proportions due to the conditions

attending
its its
deposition) should be noticed, as well as

refractive index, indicated
color
and the approximate
by the
definition of the edges either in air or in
some
denser mounting medium.

light,
The
effect
upon polarized
is
which
will
be discussed in Chapter XII,
often
of great importance.
Various peculiar phenomena
may
appear, such as hemimorphism (differences in the opposite
ends of a
crystals),
crystal),
twinning (the production of double

crystals
(parallel
skeleton
or
symmetrical
aggregates of smaller crystals), trichites (hair-like crystals,

often
more or
less
twisted),
and
sphseiulites
(radially
fibrous spherical bodies).
Sodium
and
chloride crystallized
rapidly from a solution thickened with mucilage exhibits

beautiful
skeleton
crystals;
long, curved trichites
appear in a mixture of the chlorides of chromium and

mercury.
The change
salts furnishes
in crystalline
form
of certain
compound
under the
an
interesting subject for study
microscope.
If ferrous chloride is
allowed to crystallize

144

aqueous
solution,
large, tabular,
If,
from a warm,
monoslide
symmetric (rhombic) crystals form.
now, the
be quickly heated by the application of a small flame,

the large plates
the
fall
to pieces,
and minute
crystals
(Fig.
of
anhydrous
cooling, the
salt
appear in their place
53).
On
small crystals take up water and
run
together in their original form.
Fig. 53.
Crystals of Hydrous and Anhydrous Ferrous CHLORroE. (After Lehmann.)
Behrens and other authors give a large number of

specific
microchemical
tests,
some
of
which the student
should
of
make
in order to gain practice in the recognition
typical
crystalline
forms.
Aluminium, for example,
may
be easily detected in a solution
by evaporating with
Large,
a small drop of sulphuric acid, dissolving the residue in

water,
and adding a grain
of caesium chloride.
colorless, isometric
octahedra of caesium alum are pro-
duced,
ent,
or, if
more than
1% of aluminium
sulphate be pres-
rectangular dendrites sprout from the caesium salt
(Fig. 54).
Calcium
is
best tested for
by precipitation with
sul-

MICROCHEMISTRY.
145
phuric acid, which in concentrated solution throws out

short orthorhombic crystals of anhydrous calcium sul-
FiG. 54.
Crystals of Cesium Alum.

dilute
(After
Lehmann.)
phate.
From
acid
solutions
slender monoclinic
crystallize out
prisms of the compound
(CaS04+ 2H2O)
Fig.
e,e,.
Crystals of Calcium Sulphate.

showing
(After
Lehmann.)
at C,
(Fig. 55),
numerous double twins, as
and in presence of strong acids, masses of minute needlelike crystals, as at .4.
146
3.

The Detection
earliest
of Qtiinin.
It
has been noted that
one of the
and most
profitable applications of
in the study of
microchemistry has been
made
drugs
and poisons;
extent
and the
toxicologist
depends
to
a great
upon various
method
to
specific
microchemical
tests.
Hera-
path's
for the detection of quinin, applied par-
ticularly
the
examination of the urine of patients
under quinin treatment,

It
may be
studied as an example.
crystals
depends on the formation of characteristic

the
of the iodosulphate of quinin on
addition of tinc-
ture of iodin to an alcoholic solution of the drug.

practice, the urine to be analyzed
is
In
neutralized and shaken

is
out with ether.
The
ethereal solution
evaporated and
cc.
the residue dissolved in a mixture of 12

acetic acid
of glacial
7
and 4
cc. of
95%
(i
alcohol,
to
which
drops
of dilute sulphuric acid
gram
of strong sulphuric acid
with 9 grams of water) have been added.
very minute
drop of tincture of iodin added to

duces,
if
this solution first pro-
quinin be present, a cinnamon-yellow
spot,
due
to the reaction of the iodin

little
and qumin.
Next the
alcohol separates in
drops, driving the fluid

Finally,
away
from the center of the preparation.

liquid flows
the acid
back again, and
thin, greenish plates of the
iodosulphate appe3.r, often arranged in beautiful rosette

forms.
The
crystals
produce strong polarizing
effects,
and turn brownish red when heated.

3.
The Separation
of Related Organic Substances.
The most
important aid which the microscope can offer

lies
to the chemist
in the
domain
of
organic analysis.
Lehmann even
goes so far as to say that " Crystallography
MICROCHEMISTRY.
is
I47
to the organic laboratory
what spectrum analysis

is
is
to
the
inorganic."
When
it
necessary to
separate
two
of
closely related
isomers,
to
determine the identity

diiJerent
two substances prepared by
methods, to
distinguish differences of elementary composition from
those due to allotropism, or to detect minute admixtures

of
an impurity,
it
frequently occurs that purely chemical
methods are tedious and complex, while microscopical

examination furnishes a rapid and easy solution.
general the
In
method
of study consists in
allowing the
substance in question to crystallize in contact with a body
known composition, with which it is to be compared. This may be done in three ways by adding to one
of
substance, in a melted condition or in solution, a crystal

of the other,
by preparing a mixture
from
of both in a liquid
condition and allowing the mixture to cool, and best>
perhaps,
by
crystallizing
films
of
the
two sub-
stances brought just in contact with each other.
"When
solid substances are to

is
be compared, a small
portion of one
till
it
placed under a cover-slip and heated
melts, the
amount
of material being so small as
not to reach the edge of the cover. substance

is
grain of the second
then placed just outside the cover-slip, and
in turn melted so as to flow
under and join the
first
along
(In-
line
which can be observed under the microscope.

it
cidentally
may
slight
be noted that by heating two bodies

is
simultaneously under the microscope a good measure
obtained of
the
similar
differences
of
in melting-points.
Thus
crystals
dinitrobenzol
and
dinitrotoluol
may
and
be easily distinguished.)
recrystallize, the
As two substances cool

the contact zone
phenomena along

148

and
striking.
are diverse
In some cases the mixture of
the two materials remains Hquid longer fhan the pure substances, on each side, forming a clear area in which,
later,
an amorphous precipitate
is
deposited.
The
chlo-
rides of iodin
(Fig.
56).
and
silver exhibit this case to perfection
Or a
crystalline
precipitate
different
from
a.
b.
Fig. 56.
Contact Zone between Chlorides of Silver and Iodin.

(After
Lehmann.)
either of the pure substances
may form
in the -contact
zone,
as
with silver and potassium iodides.
Or
the
crystals
may grow
across the line unchanged, forming
a single homogeneous mass.
This occurs with identical
bodies and with isomorphous substances like dibrom-
benzol and dichlorbenzol.
Or
the crystals
may grow
across the contact zone, but with spaces between them,
due
to the presence of impurities in
one or other of the
substances.
Usually the habitus of the crystals in the

will
impure
substance
be
somewhat
changed.
Pure
dinitrotoluol
and the same body containing paranitrophenomenon.

These four cases by no
toluol exhibit this
means exhaust
the possible conditions of the contact zone,
but they serve as types of some of the more striking ones,
MICROCHEMISTRY.
and may suggest how the microscope
identity
5.
sis.
149
serves to test the
and the purity
of organic bodies.
^We
The Methods
of Systematic Microchemical Analy-
have seen
that
microchemistry
is
much
wider term than crystallography, and covers the study

of change, of state
crystalline
and
In
of chemical reactions as well as
form.
tests
this
broad sense,
specific
micro-
chemical
have been suggested by Behrens and
others for the detection of the elements
and the various
acid radicles.
Recently Hinrichs has prepared a com-
plete outline for qualitative analysis

into general use
is
Whether such a scheme

doubtful, although
of time
it
will ever
come
of
promises results with a
minimum
and material; but the study
two typical basic
and acid groups may prove suggestive

an inch
in height
to the student.
A microburner yielding a flame not more

of
than a quarter
must be provided
it
for melting the
substances examined, and
is
convenient to have the
lamp
so small that
if
it
may be
placed under the stage of
the microscope
desired.
For ordinary work micro-
scopic slides are used, a drop of the substance to be tested
being placed thereon with a fine glass stirring-rod.

reagent
is
The
to
added
in
minute quantity and
is
made
mix
with the
limation
first
by means
two
of a dry stirring-rod.
For sub-
tests,
I -inch
watch-glasses are used, one
inverted over the other, with the substance to be ex-
amined
in the lower,
and the appropriate
test
reagent
borne on the concave surface of the upper,

6.
glass.
The Determination of the Metallic Elements.
Âc-
cording to the scheme prepared
by Hinrichs,
solutions of
15

first
the metallic elements are
tested
by the addition of a
fragment of metallic zinc.

silver,
is
If metallic crystals form, either

tin,
copper, bismuth, lead,
cadmium, or thallium
present.
If a blackish coating appears
upon
the zinc,
is indi-
gold, platinum, iridium, palladium, or
mercury
cated.
Should neither phenomenon follow, the other
groups of elements
nitric acid,
may
be tested for by the addition of
magnesium
ribbon,
ammonium
chloride,
and
are
magnesium and ammonium-phosphate

here,
solution;
first
we
however, concerned only with the
group of
metals, as typical of the general

If the solution contains
method
of analysis.
one of the metals precipitated

tested with several reagents,
by
zinc,
drops of
it
must be
each of which points out certain of the elements. a grain of potassium chloride
lead,
is
Thus
silver,
added, and with

is
and thallium a
characteristic reaction
obtained.
Silver salts
produce an immediate white curdy precipi-
tate of silver chloride, lead salts, crystals of lead chloride,
and
thallium
compounds, very
minute
characteristic
crystals of thallium chloride.

tests
In each case confirmatory

silver,
first
must be applied.
With
for
example, the
amorphous
precipitate should be
separated by pour-
ing off the liquid and then dissolved in a drop of
ammoon
nium
hydrate.
The
colorless solution thus obtained
warming and then cooling

silver chloride.
yields large cubical crystals of
If silver
be present, a drop of the original

faintly acid with nitric acid,
solution,
warmed and made
produces on the addition of potassium bichromate large

dark-red hexagonal plates and prisms of silver bichromate,
soluble both in nitric acid
and ammonia.
grain of
MICROCHEMISTRY.
metallic lead
151
added
to a solution of silver salts causes
the separation of metallic silver in the form of dendritic

skeleton crystals.
The
other elements are identified by similar specific
reactions.
Copper
is
at once indicated
by the
is
color of
the reduced metal
and the
solution.
Bismuth
detected
by the
large hexagonal plates of bismuth sulphate
formed
cooling.
by warming with sulphuric acid and again
Tin
solutions, with
an acid
reaction,
on the addition of
sodium
iodide,
produce characteristic crystals of sodium
iodostannites
and iodostannates.
Cadmium forms
Acid
a white
precipitate with
7.
ammonium
hydrate.
of the
The
Determination
Radicles.
In
it
studying the acid radicles in an

is first
unknown
solution,
necessary to determine the presence or absence of
organic acids.
This
may
be accomplished by mixing a
drop with a minute drop of a saturated solution of potas-
sium permanganate and adding concentrated sulphuric

acid.
In the presence of organic
acids,
nitrites,
is
sul-
phites,
and hyposulphites,
the pink solution
decolorized.
decoloriza-
The
tion
last three classes of
compounds produce
when
acetic acid is substituted for sulphuric acid,
while the organic bodies do not.

If organic acids
be absent, a drop of the solution
is
tested for volatilization with acetic acid
and sulphuric
placed on
.
acid successively.
drop of
silver nitrate
the under side of the upper watch-glass serves to absorb
the vapors
and record
their presence
by
precipitation.
in--
The
first
group of acids decomposed by acetic acid
cludes the nitrites, cyanides, carbonates, sulphites, hyposulphites,
sulphides,
and
hypochlorites.
The
second
152

by
acetic acid
group, unaffected
but volatilized by.
sul-
phuric acid, includes nitrates, acetates, chlorates, chlorides,
bromides, iodides, cyanoferrates, cyaniferrates, and
borates.
The
third
group of non-volatile acids com-
prises the sulphates, phosphates, arsenates,

(colorless
and
arsenites
solutions),
the
permanganates, bichromates,
solutions),
and chromates
each
(colored
and the
silicates,
molybdates, and tungstates (precipitates).
In
case'
specific
confirmatory
tests
are
first
made
groups
silver
for each of the acids indicated.
In the two
twice,
the
volatilization
is
repeated
once
with
nitrate
glass.
and once with lead
acetate
on the upper watchfirst
Nitrates yield negative results with the
and
Ace-
octahedral crystals of lead nitrate with the second.

tates
show the long white prisms with rhombic ends

of
silver
characteristic
acetate,
and. no
are
reaction with
the
second
reagent.
Chlorides
characterized
by
thick rhombic prisms of lead chloride

crystals
and minute cubical
of
silver
chloride.
In each case comparison

identify the
with
known substances makes it possible to compound present without serious difficulty.
REFERENCES.
Behrens, H.
by
J.
a Manual of Microchemical W. JuDD. London, 1894.
Analysis.
Trans,
CoHN, A. O.
Tests and Reagents, Chemical and Microscopical.
York, 1903. Hanausek, T. F. Lehrbuch der technischen Mikroskopie.

Stuttgart,
1 90 1.
New
HiNRicHS, C. G. Microchemical Analysis. St. Louis, 1904. Lehmann, O. Die Krystallanalyse. Leipsic, 1891. WoRMLEY, T. G. The Microchemistry of Poisons. Philadelphia, 1885.
CHAPTER
XII.
PETROGRAPHY AND METALLOGRAPHY.

I.
The Study
of
Rock Sections.The determination

more
satisfactorily carried out
of minerals can be
by the
microscopic
study of thin sections than by any other
method, and the microscope occupies almost as important

a position in the geological laboratory as in that of the
biologist.
Frequently
optical
characteristics
are
the
only ones which can be relied on for the identification of
a given mineral, and almost always they are more
easily
made
out than the chemical composition.

of minute
The
presence
and character
facility
impurities
are
detected with
under the microscope, and much information
may
be obtained with regard to the physical conditions which

attended the genesis of rock formations.
The
study of
the minute structure of fossils forms a subordinate but

attractive
branch of the general subject.

it
For examination with the microscope, and

a task of some
is
is
necessary
to prepare sections of rocks so thin as to be transparent;

this is
little
difificulty.
When
the
necessary apparatus
lathe,
at
hand,
slices are first cut
with a
using for a saw

tin
a thin disc of iron, copper, or
sheet
charged along the edge with diamond-dust

It
is,
or fine
emery.
however,
generally
possible
153
to
154
detach with a
hammer
a chip of suitable form without
using a special section-cutter.
then ground
The chip or section is down on an emery-wheel or by hand, first

glass plate with flour of emery.
is
on an iron or copper plate covered with No. 120 emery
and water, then on a
The
slice
thus prepared
cemented
to a piece of glass
about one inch square.
Canada balsam may be used
as a cement, the specimen being pressed d.own into the
warm
the
balsam, with care that bubbles are not included,

being then
firm.
slide
heated
till
the
balsam becomes
hard and
section
is
After cementing, the free surface of the

until
ground down
has
the
required
the
degree of
transparency
been attained,
glass serving as
a holder and support.

over 0.05
good section should not be
mm.
in thickness.
Certain opaque minerals are observed by reflected light,
and
in
many
cases
it
is
convenient to examine mineral

in water for
powders
directly.
They may be mounted

form
of the particles,
the study of the
and
in glycerin
or some similar highly refractive substance for internal

structure.
The
scope.
color
and form
of the crystalline or
first
amorphous
constituents of a rock are
observed under the microrelief indicate differ-
Appearances of high or low

of
refraction.
ences
The
presence
of
cleavage line3
and the occurrence
of inclusions of gaseous, liquid, or
glassy character should be noted.
The
types of crystals
present
may
be made out, care being taken to interpret

,
correctly the peculiar appearances due to the plane in
which the section studied may happen
to
lie.
Even the
15S
angle of the crystals can often be measured with a micro-
scope provided with a revolving circular stage, such as
should be used for
all
petrographical work.
The
is
inter-
section of cross-hairs placed in the eyepiece

to coincide with the vertex of
adjusted
of the
an
angle,
and one
cross-hairs
is
made to
coincide with one side of the crystal;

lies
the stage
is
then rotated until the adjacent side
along
the cross-hair and the angle of rotation measured on the
graduated edge of the stage.

definite outlines,
Incomplete crystals without
corroded crystals affected by the molten

or strained crystals, and various types
magma, broken
trographic
of incipient crystals
may
be
is
made
out.
Finally no pe-
examination
complete without the study
of polarizing properties.
it
Before going further, therefore,

its
is
necessary to describe the micropolariscope and
application.
2.
The Micropolariscope.
of
Ordinary
in
all
light
is
made
up
vibrations
of
ether
possible
directions
which he in a plane
at right angles to the direction of
transmission of the light-ray.

particle
The path
of
an individual
of ether in ordinary light
would therefore be
constantly changing.
Certain substances offer resistance
to the passage of such a light-ray, tions to
and confine
its
vibra-
two
planes at right angles to
each other.
Light
which, like that of the two resultant rays, vibrates only

in one plane,
is
known
as plane polarized light.
The
two rays produced, with vibrations in planes at right angles to each other, obey different laws of refraction.
Thick
layers of doubly refracting bodies like calcite cause
so wide a separation of the two rays as to produce two

156

re-
images of an object viewed through them (double

fraction).
The
sists,
is
polariscope
is
an instrument for detecting the

of bodies,
polarizing
power or double refraction
and conwhich
in essence, of a pair of Nicol prisms, each of
made by
cutting a
rhomb
of calcite diagonally and
cementing the halves together again, with a layer of
Fig. 57.
^Diagram of a Nicol Prism.
(After Clark.)
Canada balsam between them.

(Fig; 57)
Through such a prism

light
one of the rays of plane polarized
(EFGH),
strongly
it
known
the
as the extraordinary ray, passes unchanged, while
other
(EFKL),
first
the
ordinary ray,
is
so
refracted in the
half of the prism that
meets
the layer of
reflected
balsam
at
such an angle as to be totally

surface,
from the balsam
and
is
thus
re-
moved.
Nicol prism
is,
then, a device for
removing
I57
the ordinary ray from plane polarized light and allowing
only the extraordinary ray to pass.
For studying
is
polarization
phenomena, the microscope
provided with two Nicol prisms, properly mounted,
one placed below the stage and known as the polarizer,

the other inserted, as a rule, in the tube of the microscope
above the objective, and known as the analyzer.
One
or both of the Nicols and the stage of the microscope

are arranged to be rotated at will.
The
polarizer breaks
up
the light from the mirror into two rays of plane polarized
light,
and
of these
suppresses the ordinary ray.
The
remaining
stage
extraordinary
ray
passes
up through the
and
objective of the microscope to the analyzer.
When
the analyzer occupies two positions relative to the
polarizer, one in
which the oblique cut surfaces of the
two are
parallel,
and another
at i8o
from
this, it is as if
they were continuous, and the ray of plane polarized light

passes freely.
At the intermediate points, 90 from the no

light passes at all, since the extraor-
parallel position,
dinary ray bears to the second prism the same relation
which the ordinary ray does

suppressed.
into
to the
first,
and
is
therefore
At intermediate points the ray
is
broken up
two components, with vibrations at right angles to each other, and one of these components passes the
analyzer, producing a sort of twilight, the intensity of
parallel position is approached.
which increases as the

If,
now, the polarizer and analyzer be crossed, placed, that is, at 90 from the parallel position, no light passes
and the
of
field of the
microscope remains dark.

01
If crystals
sodium chloride
some other
salt crystaUizing in the
15S

mounted on a
slide
still
isometric system be
and placed on
remains dark.
the stage of the microscope, the stage
Conditions are unchanged, since amorphous substances

like glass
and
crystals of the isometric
system produce
no polarizing effects.
When
calcium-carbonate crystals,
or those of any other substance of the last five systems,

are examined with crossed Nicols, the crystals
visible as shining bodies
cise
become
exer-
on a black ground.
They
a polarizing action of their
own upon
the plane polarit
ized light
into
which passes through them, breaking

at right angles to
up
two rays polarized
each other and

is
not coincident with the plane in which the light

pressed
sup-
by the
analyzer.
Thus,
when viewed with

become
do
bodies
crossed Nicols, polarizing or anisotropic bodies
luminous, while non-polarizing or isotropic

not.
One
other
phenomenon must be noted

by thin
in the
most
rudimentary account of the polariscope, the production

of color effects If a plate of
plates of anisotropic substances.
mica be placed upon the stage of the microwhich we
scope, the entering ray of plane polarized light,
may
call
A,
is
resolved into two rays with vibrations at
right angles to
each other, A' and B'.
Of
is
these, the orrefracted,
dinary ray B' at right angles to
and A'
most
passes through a greater distance in the mica than does

the extraordinary ray,
and
is
retarded by an amount
depending on the thickness of the plate and the strength

of double refraction of the substance.
In the analyzer
each of the two rays A' and B'
is split
up again
into
two
components
at right angles to
each other. A' a and A'h,

B'a and B'b. A'a and B'a are
original
1^9
parallel to the plane of the
illuminating ray A,
lie
and are suppressed.
A'h
and B'h
freely.
in the plane at right angles to this
and pass
As they
unite, however, their waves, are, in differ-
ent phases, due to the retardation of B' in passing through

the mica plate,
and
if
the
amount
will
of this retardation be
right, interference
phenomena
be set
up and
very
beautiful color effects produced.
The
partieular color
varies with the thickness of the plate examined;
but at
the
two opposite positions
of the analyzer at
which most
light passes, the colors
which appear are complementary.

illustration, in
Using the notation adopted above for

case
one
A'a and B'a,
in the other,
all
A'h and B'h, pass; and,
since the
sum
of
these four rays originally produced
white
light, exist.
it is
obvious that the complementary relation
must
3.
The
Identification of Minerals.
Besides the general

it
microscopical appearance of crystals noted above,
is
possible with the polariscope to study one character of
perhaps more practical importance than any other, the

optical structure as indicated
light.
by
its
effect
on polarized
We
have seen that
crystals of the first or isometric
system are isotropic.
All other crystals are anisotropic,
but in those belonging to the tetragonal and hexagonal

systems there
crystals is
is
one axis about which the structure of the

this axis light passes
homogeneous, and along
unaffected.
Such
crystals
are
called
uniaxial,
while
crystals of the orthorhombic, monoclinic,
and
triclinic sys-
tems possess two such axes, and are called
biaxial.
it
Examination with crossed Nicols makes
possible at
loo
ELEMENTS OF /tPPLIED MICROSCOPY.
once to distinguish isometric crystals from those of other

systems.
Good
crystals to
compare
in this respect are
garnet and gypsum, the former being isotropic, the latter

anisotropic.
The same
minerals furnish an
instructive
contrast in refractive index, the former showing high
and
the latter low reUef.
Tetragonal, 'hexagonal, and orthorhombic crystals
may
tri-
be separated from those

clinic types
of
the monoclinic
and
by the
fact that in
them
the directions of
vibration (sometimes called axes of elasticity) are parallel to
the crystal axes of the crystal.
The
position of
these vibration directions can be determined
by noting
then that
the position in which the crystal becomes dark (position

of extinction)
with crossed Nicols, for
it
is
the vibration directions coincide with the planes of the

Nicols.
cross-wire placed in the eyepiece so as to
coincide with the plane of vibration of the analyzer under

these conditions coincides also with the axis of elasticity
of
the
crystal.
By now removing
the
analyzer,
the
cleavage lines and boundaries of the
crystal
may be
seen and the stage rotated so that the cross-wire corre-
sponds with them.
The
angle of rotation required to
produce
this effect as
if
measured on the graduated edge of
the stage,
o or 90, on all the crystals examined, indi-
cates a tetragonal, hexagonal, or orthorhombic crystal.
Such substances are said

or to be symmetrical.
condition, while in
to
show no
extinction angles
illustrates this
all
Quartz (hexagonal)
(monoclinic)
angles.
Gypsum
extinction
planes but
like
one
show
large
This,
other
monoclinic minerals, exhibits in one plane phenomena

similar to those characteristic of hexagonal crystals,
i6l
and
shows no extinction angle.
When
certain sections of anisotropic crystals are
exam-
ined with convergent polarized hght obtained by a converging lens fitted over the polarizer, interference figures
are produced, consisting of dark or colored rings
crosses.
It is
and
impossible to enter here into the optical
principles
which condition these phenomena;
but the
difference between the circular figures produced axial crystals,
by
uni-
and the
elliptical
appearances characteristic
Fig. 58.
Intekfeeence Figukes.
(After Luquer.)
of biaxial crystals (Fig. 58), are easily apparent.
With
some
sections
which show only an

if
indistinct
bar instead
of a clear figure,
it
the stage be rotated, the bar follows
and remains
straight, in the case of uniaxial crystals,

it
while with biaxial crystals

tion
moves
in the opposite direcwill again
and becomes curved. Quartz and gypsum
serve for comparison, the former being uniaxial, the latter

biaxial.
Two more
characters remain to be noted, absorption
62

Absorption
is
and pleochroism.
all
the
power
of absorbing
or part of the light which vibrates in certain planes,

similar property of removing the
If a rock
and pleochroism the

section be
rays of certain colors in particular planes.
examined with
parallel rays of polarized light
and the
tion will
stage be rotated, minerals which exhibit absorp-
show a change
in intensity of light
from the
usual shade almost to black, while pleochroic minerals
show a change
in color, or at least in shade.

sufficient to identify
We
six
have
now data
any one
of the
crystallographic
systems, whose
optical
characters
may
gent
be briefly summarized as follows: Isometric system;
isotropic, exhibiting
light.
no interference
figures in converuniaxial,
inter-
Tetragonal system:
symmetrical,
sections
anisotropic,
extinction
giving
uniaxial
ference figures, often with rectangular cleavage.
Hex-
agonal system: anisotropic, uniaxial, extinction symmetrical,
sections giving uniaxial interference figures, three-
or six-sided, or
of 60.
tinction
show cleavage
lines intersecting at angle
Orthorhombic system: anisotropic,

symmetrical in
all
biaxial,
exsys-
sections.
Monoclinic
tem:
anisotropic, biaxial, extinction symmetrical in one
zone, while other sections

clinic
all
show
extinction angles.
Tri-
system:
anisotropic, biaxial, extinction angles in
sections.
By
application
of
of
these
criteria,
comoptical
bined
with
observation
the
more obvious
it is
characters, color, aggregation, etc.,
possible to identify
certainty.
any of the commoner minerals with ease and

4.
The Structure and Behavior
of
Alloys.
Metal-
lography, or the study of the structure of metals with the

microscope, owes
its
163
beginnings to Dr. Early of Sheffield

of Charlottenburg.
Its
and Professor Martens
aim
is
to
study the minute structure of metals with the microscope,

not, as in the case of minerals,
by grinding off thin sections,

light.
but by examining polished surfaces by reflected

Its
importance
steel as to
is now so great in demand some attention
the testing of iron

in the
and
most elementary
it is
survey of the
field of
microscopy; and since

is
with the
various alloys that metallography
principally 'concerned,
certain general characteristics of these bodies
must
first
be
briefly discussed.
Many alloys may,

and copper
is
for practical purposes, be considered

silver
as solutions of one metal in another; the alloy of
a good example of this type.
If
a molten
alloy containing
more than 72%

is
of silver
be gradually
cooled, a point
reached at which the rate of cooling

silver
becomes retarded and

greater the
ture at
begins to separate.
is
The
when
silver
amount
of silver the higher
the tempera-
which
this process begins.
After a time,
the
still
the temperature falls to 770

will
C,
molten
be found to have fallen to just 72%; a second retardais
tion in the cooling
now
apparent accompanied by the

If
solidification of the entire alloy.
more than 28%

first
of
copper be present, on the other hand, the

is
retardation
accomplished by a deposition of copper, which con-
tinues until the excess has all been solidified, which occurs
at
a temperature of 770 C.
The remainder then solidifies
entire.
silver
tion,
molten alloy containing originally
72%
of
and 28%
of copper shows only one point of retarda-
remaining molten above 770 and becoming solid
164
ELEMENTS OF
below that temperature.

points
is
The
relation of these various
made more
clear in Fig. 59, in
which the outer
limits of the oblique lines represent the fusing-points of
pure silver and copper.
Silver
100
PETROGRAPHY AND MET/tLLOGRAPHY.

5.
165
Metallography
of
Steel.
Steel
is
practically
an
alloy of ferric carbide
and
iron,
and the phenomena attend-
ing
the
its
cooling from a high temperature are essentially

just considered.
same as those we have
The com-
pound corresponding
FejC, or
to the eutectic alloy contains
12%
0.8%
of pure carbon.
This substance
is
known
as pearlyte,
and
steel of the right
composition on gradual
cooling from a fused condition becomes entirely con-
verted into
it
at
a temperature of 670 C.
is
If
more
car-
bon be
present, ferric carbide

if
separated before this
temperature be reached;
iron, that
metal
is
segregated.
The
up
carbide
is
known
to microscopists as cementite, the
pure iron as
ferrite.
Very low carbon
steels are
made
of ferrite, with here
and there
particles of pearlyte.
The
proportion of pearlyte increases with an increase of
carbon up to 0.8%, after that point being mixed with

cementite.
If steel
be suddenly cooled from a high temperature,
above that at which segregation begins, a new com-
pound
stance
is is
formed known as martensite, and due the intense hardness

at
to this sub-
of steel thus quenched.
Below the point
which
ferrite or
cementite begins to
separate, sudden cooling produces a mixture of these
bodies with martensite.

If a
smooth surface
of steel
be examined under the micro-
scope the presence and the general proportions of these

various
compounds may be
readily
size,
made
thick,
is
out.
The
very
specimen, of some- convenient

to
say from half an inch

first
an inch square and half an inch
carefully polished
upon one
tace with
emery and with
66
rouge.
The
final polishings are carried out
by rubbing
the metal on a layer of parchment stretched over smooth
wood and covered with moist rouge

ammonia-water has been added.
the softer parts are
structures stand out in low relief.
finally
to
which a
little
In
this latter process
somewhat eroded and
the harder
The specimen when
prepared
is
examined with an ordinary micro-
scope provided only with some apparatus for illuminating with reflected
light.
This
may
consist merely of a con-
denser or reflector which throws light obliquely on the

stage, or better, of
fitting into the
a plane reflector or right-angled prism
tube of the microscope above the objective.

its
This
is
perforated at
center to allow passage of rays

its
upward from
reflects light
the object, while the remainder of
surface
the
from without
vertically
downward on
specimen.
When examined
pearlyte
is
in this fashion with the high objective,
seen to be
made up
of very
minute
crystals of
two substances arranged alternately;

teristic structure of all eutectic alloys,
this is the charac-
and
indicates that
in spite of their constancy of composition these bodies are
merely peculiarly
stituent substances.
intimate
mixtures
if
of
the
two conappears in
it
Cementite,
present,
large
whitish
masses;
relief.
being harder than pearlyte,
stands out in
Ferrite,
;
on the other hand,

alloys with
is
the
softest material in steel
and in
more than 88%
of free iron pearlyte areas
appear standing out from a
background of
of
ferrite.
Fig. 60 illustrates the structure

as
mixed pearlyte
If
and cementite
seen -under the

it
microscope.
martensite be present
appears as a

homogeneous substance made up
easily distinguished
167
of
ciystalline needles
from any other constituents

of carbon in steel
of steel.
deter-
The
exact
amount
must be
Fic:. 60.
Microscopic Appearance as Steel with j..5% Carbon (Peaelytf and Cementite). (After Sauveur.)
mined by chemical methods; but the presence of the mineral forms we have discussed and the manner in
which they are associated can only be ascertained under
the microscope.
Furthermore, the crystals present

arranged,
to
may
be
abnormally
being
perhaps
not
closely
packed or so large as
produce incipient cleavage planes.
68
All these factors are of great importance in determining
the hardness of steel;
it
is
therefore natural that the

in
microscope should have become indispensable

laboratory of the iron and steel expert.
the
Its practical
application to the study of other alloys promises extensive
development in the future.
REFERENCES.
Clark, C. H.
1894.
Practical
Methods
in
Microscopy.
Boston,
LuQUER, L. M. Minerals in Rock Sections. New York, 1898. Zerkel, F. Lehrbuch der Petrographie. Leipsic, 1893. RosENBUCH, H. Iddings, J. P. (translator). Microscopical Physiography of the Rock -making Minerals. New York,
1893.
HiORNS, A. H. Metallography. London, 1902. Sauveur, a. The Constitution of Steel considered as an Alloy of Iron and Carbon. Technology Quarterly, XI, 1898, 78.
INDEX.
PAGB
Abbe
"
condenser
I9i 27, 12, 14, 35,
19
28 36
Aberration, chromatic
"
spherical
12, 14, 35,
36
152
Acetates, determination of
Achromatic objective
(see Objective).
Acid hsematin " radicles, detection of

Acids, detection of
134
151
151
Adjustable objective
32
Adjustments (see Microscope, adjustments). Adulterants of drugs (see Drugs).
"
"
foods (see Foods).
Agave
Agglutinins
Air-bubbles
89
119
30
49
,
Albumin fixative Aluminium, test for

Alcohol, as a fixing agent
144
44, 45
Alhazen
AUotropism.
Alloys, behavior of
,
10
147 162 163
"
nature of
Alpaca
Alterations in manuscripts
91
138
iSi 17
Amici
Ammonium-magnesium phosphate Anabana in water
in urine
107 123
169
70
INDEX.
PAGE
Aniemia Analyzer
Angiosperms,
fibres of
113
157 loi
5
Angle of prisms Angular aperture
17
Animal
hairs (see Wool).
Anisotropic bodies
158
135
17
17
Anthropometry
Aperture, angular
"
.numerical
Aplanatic lenses (see Objective).
Arabian microscopy
lo 9 10
Aristophanes
Armati
Arrowroot
Asbestos
Asterionella in water
66
81
124.
Axial illumination
Axis, principal
26
6
I17
116
;
Bacillus diphtheri(B
^ V
<
tuberculosis
Bacon
Balsam
(see
10 84
142, 149
Mounting Media, balsam).
Bast-fibre
Behrens Bertillon
Biaxial crystals
135
1
59
Biconvex lens (see Convex Lens).

Birch fibres
loi 151
Bismuth, microscopical tests for

Blood, examination of cells
109
1
"
' '
"
for malarial parasites
14
spectroscopy of
Blood-cells
'
133 Ill 112

107, 108
'
counting
in urine
"
"
"
normal numbers
staining
113
Ill
Blood-smear, preparation
Blood-stains, detection of
109 126
1 I
INDEX.
'page
Blood-Stains, microscopical test
127
"
Baehmeria.
microspectroscopical
,
128
88
Bombyx
Basophiles ....
g2
iii.
Brownian movement
Buckwheat-starch
Burning-glass
37 78 9
tests for
Cadmium, microchemical
Gsesium alum, crystals of
IJ
144
107
145
._
Calcium oxalate in urine " sulphate crystals

".
test for
144
92
54. 5Si 5^
55,
Camel's hair
Camera Lucida " " construction of " " lighting with " " measurement with " " use.of Canada balsam (see Mounting Media, balsam).
.
, ,
56
55
54 56 86
116
167 91
Cannabis
Carbol-fuchsin
Carbon in Cashmere
steel
Cassavas (see Sago).

Cell theory
.'
105
Cells (see Mcûnting, cells).
Cellular pathology
105
165, 166
Cementite
Chevalier
'.
15
Chicory (see Coffee, adulterants).

Chlorides, detection of
,
152
Chromatic aberration (see Aberration). Chromium and mercury chloride

Clearing agents
Coffee adulterants
143
45
,
>
4^
73 7^
"
microscopic structure
Color production by mica plate
158 i
Condenser
(see
Abb
Condenser).
Coniferae, fibres
172
Index.
PAGE
Contact zone
147 132
Continuous spectra
Convex "
' '
lens, formation of
images
4,
6 6
163
151
"
Refraction
by
tests for
Copper, alloy with silver

,
microchemical
Corchorus
Corn-starch
59,
,
87
64
65 81
"
Cotton
microscopic structure
" "
" "
-fibre
81,
82
82 83
" "
"
canal
chemical treatment
composition
mercerization
8z
"82
"
" "
" "
.length
,
" preparation Cotton paper

;
83 82
95
31,
Cover-glass, effect of
32
41 143
"
measurement of
32, 39
Cover-glasses, cleaning
Crystal angles
"
"
measurement
Cryohydrate
Crystals (see Minerals).
155 164
"
.anisotropic.
.biaxial
,
159.161,162
159 142
*.
" " "
formation of
"
"
'
'
.habitus , hexagonal
, ,
143
159, 162
159, 160, 162
isotropic
microscopic study of
142
159, 160. 162 159, 160, 162
"
"
monoclinic
orthorhombic
tetragonal
triclinic
"
"
"
159, 160. 162

159, 160. 162
uniaxial
159 6
Curvature of lens
"
Cytology
,
" the
. . .
field
35 105
27
Dark-ground illumination
INbEX.
173
PAGE
Dehydration, in glycerin " with alcohol

,
43
45 10 117
Descartes
"
Diagnosis of Diphtheria
"
*
'
" Tuberculosis " Typhoid Fever

(see Microscope,
,
116 118
Diaphragm
Diatoms
diaphragm).
28
35, 124
use of
Differential blood count
112
49, 50
"
staining
Dinitrobenzol
Dinitrotoluol
147 147
Diphtheria, diagnosis of
117
137
Documents, study of
Double staining Draw-tube, use of
50
20, 31, 32
II
Drebbel Drug adulterants

Dry mounting
Drugs, microscopically examined for adulterants (see Mounting Media, air).
69 69
Early Ehrenberg
Eosin and hsematin, double staining
163
106
50
11
Eosinophils
Epithelial cells in urine
109
iii 103
Erythrocytes
Esparto-fibre
Eutectic alloy
164
2
Eye, structure
Eyepiece (see Microscope, lenses).

Eye-point
Eyes, care of
Fabrics, examination of
'.
33 33
94
165
165, 166
Ferric carbide in steel

Ferrite Fibres, analytical
key
81,
" "
animal
effect of reagents
98 90
81
174
INDEX.
PAGE 80
Fibres, kinds of
" " "
.textile
,
80
81
vegetable
wood-pulp
100
102 137 136
,
Fibrovascular bundles
Finger-print classification
'
'
patterns
Finger-prints
Fir-fibre
135
100
Fixative,
Mayer's albumin
.-
49
Fixing
''
agents
Flax " -fibre
44 44 84
85
Flemming's Mixture
Focussing
Focus, principal
44
29 6
Food adulterants
Forgery, detection
of.
69
138
15
Fraunhofer
"
lines
132
Freezing microtome
49
11
Galileo
Galton Germ theory
of disease
135 106
jelly).
Glycerin (see Mounting Media, glycerin).

Glycerin jelly (see Mounting Media, glycerin
Goring
Gossypium
....,,.
(see Trichina).
15
81
Government inspection of meat
Gower's solution Gymnosperms, fibres
112
100
,
Gypsum
Hsemalum, Mayer's
161
50
114
Hamamteba
Hsematin and
eosin, double staining
50
134 112
,
HEematin, spectrum of
Hsemocytometer
Haemoglobin, spectra of
134
INDEX.
175
PAGE
Handwriting " -lines, microscopic appearance
138
of.
140
"
statistical
study
of.
139
142
;
Harting
Hemimorphism
Hemlock-fibre
143
100 86
86
Hemp
"
-fibre
,
" "
Manila (see Manila Hemp).

Sisal (see Sisal
Hemp).
135 105
Herschel
Hertvvig HiNRICHS Homogeneous immersion
149
16
HooKE
Huyghenian ocular
Illuminating power
104
22
35 8
i
Illumination (see Lighting).
Image, construction of " , definition of " ., formation of by object inside principal focus " , formation of by object outside principal focus " real
, ' ' ' ' ,
7,
7,
7,
8
8
i
7,
retinal
retinal, formation of
"
' '
retinal, interpretation of
virtual
7,9
II, 13
Images, formation in compound microscope formation of, by convex lens

' '
Imbedding in " "
celloidin
49
47
paraffin
Immersion, objective (see Objective). Infusoria in water (see Water, Drinking-).

Ink, examination of
Interference effects
137
IS^
Interpretation of appearances
36
2
"
"
retinal
image
lodin chloride, crystals
148
lodo sulphate of quinin

Iron in steel
146
i^S
176
INDEX.
PAGE
Isomers, separation of
Isotropic bodies
147
158
Janssen...
Jute
II
87
-fibre
"
87
10
105
KiRCHER KOLLIKER
Larch-fibre
loo
tests for
10, 104,
Lead, microcheinical
150
106
Leeuwenhoek Lehmann
Leishman's blood-stain
Lens, curvature of
142, 146
51; iii, 114
6
iii
Leucocytes, various types

Leucocytosis
113
25, 26, 27, 28
Lighting
Linen paper Line spectra I.inum
95 132
84
15
Lister Loeffler's methylene blue Lymphocytes

Magnification, measurement of
117 112
57
Magnifying power
Malarial parasite
34
114
Malpighi Manila hemp " paper
104
89
97
163
1
Martens
Martensite
66
Massachusetts State Board of Health
71, 122
Mayer's albumin fixative hsemalum Meat inspection (see Trichina).

'
'
49 50
Mechanical stage
Mercerization
Metals, microchemical tests for.
.
19
83
. , ,
,
1^0
INDEX.
177
Metals, separation of in alloys

Metallic elements, determination of
163
Metallography " of
149 162
165
steel
Methylene blue Microchemical analysis, apparatus " " of acids
117
for
149
I c I
" "
"
'
"
" "
tests
,
" metals
possibilities of
,
149
142
systematic
149 144
'
Micrometer, measurement with " ocular

,
54
53 52 53
155, 157
.stage
"
standardization of
Micropulariscope
Microscope, adjustments
' '
20
20
23
10, II, 12, 13
arm
cleaning
" "
'
'
'
'
compound diaphragm draw -tube

function of
history of
-
19
20
3
,
"
' '
"
"
Hooke's
biology
9 12
in
104
" "
" "
"
' '
in medicine
,
and sanitary science
105
II
20, 21
Leeuwenhoek's
lenses
"
,
cleaning of
24
57
17
magnification of
mechanical parts
mirror
pillar
"
19 19
"
"
position of
qualifications of
23
"
"
34
23
setting
up
simple
9
19
17
" "
f
.stage
stand
tube
,..,,..,,.,
20
178
INDEX.
PAGE
Microscopy, forensic
126
',
Microspectroscope
130
128
48, 49
Microspectroscopy of blood-stains
Microtome
Minerals, identification
in rock sections
159162
154
Minot-Blake microtome
Mohair Motion under the microscope Mounting cells Mounting media, air " " .balsam "
48
qi
36
41,
42
41
43
" "
" "
" "
"
,effectof
,
38
42 43
38,
jelly---.
gJycerin
"
" "
"
refractive effects
requisites of
39 40 39
temporary
Musa
Muscce
volit antes
,
89
34
75
Mustard, adulterants
"
microscopic structure of
74
22
Negative ocular
Neutrophiles
Nicol prisms
Nitrates, detection of
112
156, 157
152
i
Normal
vision
Numerical aperture
Objective (see Microscope, lenses).
17
" " "
achromatic
aplanatie
14, 15
16
15, 16
immersion
Oblique, illumination
27
Ocular (see Microscope, lenses).

Oil-drops
Olive-stones (see Pepper).
30
15
Organic acids, detection of " substances, examination of

Overstaining
146
50
INDEX.
79
PAGE Ovis
gl
Oxyhaemoglobin, spectrum of Oxyphiles
134
m
95,
Paper and paper making " -fibres, analytical key " manufacture
' ' ' ' ,
96
98
96 96
97 95
raw materials
and amounts
stock, kinds
Papyrus
Paraffin
imbedding
,
47
148
95
Paranitrotoluol crystals
Parchment
Pasteur
Pearlyte in steel
106
165
Pedesis
Penetration
37
35
Pepper, adulterants
78
"
microscopic structure of
76
153
Petrography
Phloem
Piper
(see Bast-fibre).
7&
155 158
9 155, 156
Plano-convex lens (see Convex Lens).
Plane polarized light

Pleochroism
Pliny
Polariscope
"
structure of
156
157 155
Polarization with microscope
Polarized light
Polarizer
157
>
Polarizing microscope
Poplar-fibres
157 loi
Pork, examination for Trichina (see Trichina).
Potassium iodide crystals

Potato-starch
148
59, 61
"
microscopy of
63 6 6
5
Principal axis
"
' '
focus
Prisms, effect of angle

,
refraction
by
i8o
INDEX.
PAGE
Qualitative analysis, microchemical
14.9
Quartz
Quinin, test for
l6i
146
,
Ramie
"
-fibre
88 88
Real image (see Image).
Rkaumur
Reduced hsemoglobin
Reflected light
95 134
28
Refraction by convex lens
4
3,
" "
"
plane surfaces
4
5
" prisms
,
"
"
Retina
laws of
toward normal
4
35 I, 2
2,
Resolving power
Retinal image
Retting
Rice-starch
84 66
154
153
153
Rock " "

Sago
sections, characteristics of
"
preparation of
"
study of
schleiden
67 104
104
Schwann
Scutching
Section cutting
84 46
"
"
"
by hand
46
47
"
"
"
microtome..,,
" in pith Sedgwick-Rafter method

Selligues
46,47
122
15
Seneca Serum reaction

"
Silk,
test for
9 118
Iig 92
93 163
typhoid fever
formation of
structure of
"
Silver, alloy
with copper
"
chloride crystals
148
,
"
iodide crystals
148
INDEX.
i8i
PAGE 150
89 143
Silver,
microchemical
tests for
Sisal
hemp
Skeleton crystals
Slides, cleaning
41
143
Sodium
"
chloride, crystals
Spectra of gases and solids
132
. :
" organic bodies.
133
131
Spectroscope
Sphserulites
143
Spherical aberration (see Aberration).

Spruce-fibre
100
115
Sputum, examination of
Staining
49
,
"
blood-cells
ill
" " "
diphtheria bacilli
tubercle bacilli
117
116
with hsematin and eosin
50
Stage micrometer (see Micrometer). " (see Microscope, stage).

Starch, as index of food adulterants
"
' '
in baking
powder
69 62
" " " " " " " "
commercial uses for conversion into sugar

formula of
in chlorophyll
60
62
5^
in flour in grains in potato
59 61
60
59, 61
microscopy of
nature of
origin of

62
5^ 5 61 59
" " "
paste
;
preparation for market

prices of
refining, of
"
" "
^
59 i
uses of
Starches of pea and bean

State Board of Health of Massachusetts, work on food adulterants.
Steel,
66
71
165 165
composition of
"
metallography of
Steno, law of
143
INDEX.
PAGE
Stipa
...,,.
loi
oy
103
IO4.
Straw-board
Straw-fibre
SWAMMERDAM
Synura
Tapioca
in water
(see Sago).
124
Telescope, function of
3
tests for
Thallium, microscopical
150
Thoma
microtome
48
151
Tin, microscopical tests for
Toxicology
146 100 120
Tracheids
Trichina
Trichinosis (see Trichina).
Trichites
143
Tube
"
casts in urine
log
length, standard
bacilli
1
31
16
Tubercle
Tuberculosis, diagnosis of
116
15
TULLEY
Turntable
42
143
Twinning Typhoid bacilli, agglutination of " fever, diagnosis of

Ultra-violet light
:
119
1
18
21
Uniaxial crystals. ......
159
107
Urates in urine
Uric acid in urine
107 107 107
Urinary sediment
Urine, examination of
Uroglena in water
124
105 105
Van Beneden
ViRCHOw
Virtual image (see Image).
Vision, normal
Walker
Water, microscopy of drinking
129
121
122
"
filtration of.
INDEX.
183
PAGE 64 64
97
1
Wheat-starch " microscopic structure

,
Whitney WiDAL reaction Wilder Wiley Wood pulp

"
19
135
60
97 ICO
",
"
fibres.. ....
paper
95,97
97
;
Woodman
Wool, kinds and qualities
' '
92
90
51, iii,
characteristics of
Wright,
blood-stain
114
137
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Elements of Applied Microscopy. A Text-Book For Beginners (1905) by Winslow

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Elements of Applied Microscopy. A Text-Book For Beginners (1905) by Winslow

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CORNELL

Cornell University Library

3 1924 003 015 751

Cornell University Library

Cornell University Library.

There are no known copyright

the United States on the use of the

CHARLES-EDWARD AMORY WINSLOW,

Massachusetts Institute of Technology.

JOHN WILEY & SONS. London: CHAPMAN & HALL, Limited.

CHARLES-EDWARD AMORY WINSLOW.

ROBERT DRUMMOND, PRINTER, NEW YORK,

'Elements of Applied- Microscopy.

book for Beginiiers.

Massachusetts Instiobject of the course

give facility in the manipulation of the

four chapters consider the micro-

and the other eight

deal with the starches, adulterations

ft food and drugs, textile

inicrochemistry, petrology and metallurgy.

to give the student the principles

the point of view.

Exercises are then given

methods necessary for the

elucidation of the questions which arise, in

and was designed

for use by a teacher pos-

DEDICATED BY THE AUTHOR Zo 1bl6 ^otber.

intended for the teacher, and the

beginner with the microscope, not for the

the branches of technical microscopy have

been already made the basis for monographs with which

other hand, there are

entertaining popular books

on the microscope which

Neither type of work was

suited for the use of a class in Industrial Microscopy

offered to second-year Chemists

object of this course

to give facility in to fur-

the manipulation of the microscope;

nish an acquaintance with the scope of

which should take up the fundaitself

mentals of the science and art of microscopy

a rapid but wide survey of the principal fields in

which the microscope has been applied

such elementary but comprehensive work

Enghsh among the numerous able

special branches of the subject.

therefore the outgrowth of a pedagogic need.

from the standpoint

expert in any of the

conveys to the student's mind such an idea of the possible

eventually to the further exploration

quoted in connection with the various chapters have been

further desires to ex-

press his grateful obligation, to Dr. P. G. Stiles for the

preparation of original drawings;

assistance in regard to various portions of the manuscript

Acknowledgments are due

been copied directly or redrawn

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