MICROSCOPE

A microscope is an optical instrument which is used to see highly

magnified images of tiny objects such as bacteria, cells , viruses and
protozoans etc. because these tiny objects cannot be seen by naked eyes.

Simple Microscope

A simple microscope is also called magnifying glass. It is actually a

convex lens of small focal length, which is used for seeing the magnified
images of small objects.

Principle of Simple Microscope

A simple microscope works on the principle that when a tiny object is

placed within its focus, a virtual, erect and magnified image of the object
is formed at the least distance of distinct vision from the eye held close to
the lens.

Magnifying Power of a Simple Microscope

The magnifying power or angular magnification of a microscope may be
defined as the ratio of the angle subtended at the eye by the image
formed at the distance of the distinct vision to the angle subtended by the
object when placed at the distance of the distinct vision.
The ray diagram shows that the image of the object AB is formed at A1B1.
A1B1 is formed at the least distance of distinct vision.

The figure shows that the angle A1OB1 subtended at the eye by the object
in the position A1B1 is greater than the angle AOB subtended by it in the
position AB. From this it is clear that the eye estimates the angle
subtended by an object on it and not the linear size of the object.
But OB1 = Least distance of distinct vision from the lens or eye = D
OB = u = distance between the lens and the object

The distance between the image and the lens is negative as the image is
virtual.

The lens formula for a convex lens is

Where f is the focal length of the lens

Multiplying both sides of the equation (1) by v we get

In the case of a simple microscope v = -D

D = 25 cm

From equation (3) it is clear that a convex lens of short focal length has a
large magnifying power.
The highest magnification which can be obtained from a simple
microscope is about 20.

Uses of Simple Microscope

Following are the important uses of simple microscope:

1. The simple microscope is commonly used by watch makers to see
the magnified view of small parts of a watch.
2. It is also used by the jewelers to see the magnified view of the fine
parts of jewellery.
3. Simple microscope is used to see the enlarged image of letters of a
book, textures of fibers or threads of a cloth.
4. Simple microscope is used to see the magnified view of different
particles of different types of soils.
5. It is used by palmists to see enlarged view of the lines of our hand.
6. Simple microscope is used by skin specialists to find out various
diseases of skin.
7. It is also used to see the details of stamp and engravings.

Slide Mount Instructions

Before you start building your slides, make sure you have
everything you will need, including slides, cover slips, droppers or
pipets and any chemicals or stains you plan to use.
You will be using two main types of slides, 1) the common flat glass
slide, and 2) the depression or well slides. Well slides have a small
well, or indentation, in the center to hold a drop of water or liquid
substance. They are more expensive and usually used without a
cover slip.
Standard slides can be either plastic or glass and are 1 x 3 inches
(25 x 75 mm) in size and 1 to 1.2 mm thick.
Wet slides will use a cover slip or cover glass, a very thin square
piece of glass (or plastic) that is placed over the sample drop.
Without the cover in place, surface tension would cause the droplet
to bunch up in a dome. The cover breaks this tension, flattening the
sample and allowing very close inspection with minimal focusing.
The cover also serves to protect the objective lens from interfering
with the sample drop.

MOUNTS
There are four common ways to mount a microscope slide as
described below:

Dry Mount
In a dry mount, the specimen is placed directly on the slide. A cover
slip may be used to keep the specimen in place and to help protect
the objective lens. Dry mounts are suitable for specimens such as
samples of pollen, hair, feathers or plant materials.

Wet Mount
In a wet mount, a drop of water is used to suspend the specimen
between the slide and cover slip. Place a sample on the slide. Using
a pipette, place a drop of water on the specimen. Then place on
edge of the cover slip over the sample and carefully lower the cover
slip into place using a toothpick or equivalent. This method will help
prevent air bubbles from being trapped under the cover slip.

Your objective is to have sufficient water to fill the space between

cover slip and slide. If there is too much water, the cover slip will
slide around. Take a piece of paper towel and hold it close to one
edge of the cover slip. This will draw out some water. If too dry, add
a drop of water beside the cover slip. Practice this until you get used
to it.
Wet mounts are suitable for studying water-bound organisms such
as paramecium or bodily fluids such as saliva, blood and urine.

Section Mount
In a section mount, an extremely thin cross-section of a specimen is
used. Using a microtome, cut a thin slice of your selected specimen
such as an onion, and carefully set it on your slide. Then follow the
instructions for a dry or wet mount. A stain can often be applied
directly to the specimen before covering with a cover slip.
Section mounts are suitable for useful for a wide variety of samples
such as fruit, vegetables and other solids that can be cut into small
slices.

Smear
A smear is made by carefully smearing a thin layer of the specimen
across a slide and then applying a cover slip. Typically, a smear
should be allowed to air dry before applying a stain.

STAINS
Stains are used to help identify different types of cells using light
microscopes. They give the image more contrast and allow cells to
be classified according to their shape (morphology). By using a
variety of different stains, you can selectively stain different areas
such as a cell wall, nucleus, or the entire cell. Stains can also help
differentiate between living or dead cells.
Stains tend to be grouped as neutral, acidic or basic, depending
upon their chemical makeup and will attract or repel different
organisms accordingly. For example, scientists and health
professionals use Methylene Blue, a slightly alkaline stain, to reveal
the presence of deoxyribonucleic acid, more commonly known as
DNA.
 Stain Types
Iodine is one of the more commonly available stains and is used to
identify starch in a variety of samples. It will stain carbohydrates in
plants and animal specimens brown or blue-black. Glycogen will
show as red.
Methylene Blue is an alkaline stain useful in identifying acidic cell
nuclei and DNA in animal, bacteria or blood samples. Its also useful
in aquariums to prevent the spread of fungal infections in fish.
Eosin Y is an acidic stain which stains pink for alkaline cells
(cytoplasm, for example). It colors red for blood cells, cytoplasm and
cell membranes. Eosin's most important medical uses are in blood
and bone-marrow testing, including the PAP smear.
Gram's Stain is one of the most frequently used processes in
identifying bacteria used daily in hospitals. It is a primary test that
quickly and cost effectively divides bacteria into one of two types:
Gram positive or Gram negative.

STAINING STEPS
1. Prepare a wet mount slide.
2. Collect a drop of stain with an eye dropper or pipette.
3. Put a drop of stain on an outer edge of your cover slide.
4. Place a piece of napkin or paper towel against the opposite side of
your cover slip, right up against the edge. This will help draw the
stain under the cover and across the specimen.
5. You may need to add another drop to ensure complete coverage.
6. The slide is now ready for viewing.
Pay attention to the following do's and don'ts:

1. The Do's:
a. Wash your hands before handling slides to avoid transferring
finger grease to the slides.
b. To remove oil from an objective, moisten a piece of lens paper
with ethyl ether and slowly draw the wet paper across the front
surface of the lens.
c. To remove dirt from an eyepiece, breathe on the lens and
GENTLY wipe off the condensed water with lens paper.
d. Get help if these steps do not solve your problem.

2. The Don'ts:
a. Never touch a lens with anything but lens paper.
b. Never touch a lens with dry lens paper.
c. Never rub a lens.
d. Never put oil on a dry objective, and remove it at once if an
accident occurs.
e. Never wear mascara when using a microscope.