Stains/ Dyes called the Mallory staining technique uses

three acidic dyes: aniline blue, acid fuschin

STAINS/ DYES and orange G, which selectively stain
They are colored organic compounds used
for staining microorganisms. Chemically,
collagen, cytoplasm and red blood cells
Stains= Benzene ring+ Chromophore+
Auxochrome Basic dyes

According to nature of stain, it can be classified Methyl green Green

into: Methylene
blue Blue
1. Acidic Dyes: It is dye which has negative
charge so they bind to positively charged cell Pyronin G Red
structures like some proteins. Acidic dyes are Toluidine blue Blue
not very often used in Microbiology lab.except to
provide background staining like Capsule Acidic dyes
staining. Examples: Nigrosine, Picric acid,
Eosin, Acid fuschin, India ink etc. Acid fuschin Red

2. Basic Dyes: This dye have positive charge & Aniline Blue Blue
bind to negatively charged molecules(nucleic
Eosin Red
acid, -COOH -OH). Since, surface of bacterial
cells are negatively charged(due to Teichoic Orange G Orange
acid), basic dyes are most commonly used in
bacteriology. Examples: Crystal Violet,
Methylene Blue, Safranin , basic fuschin. respectively.
3. Neutral Dyes: They are usually formed from
precipitation in which are produced when
Periodic acid-Schiff
aqueous acidic & basic stains are combined.
Neutral dyes stains nucleic acids, & cytoplasm. reaction (PAS)
Eg; Eosinate of Methylene blue, Giesma stain.
The Schiff reagent is a bleached basic
fuschin that reacts with aldehyde groups.
Histological Stains This reaction results in a deep red colour in
the section. It is the basis of the PAS stain.
other than H&E PAS stains carbohydrates and carbohydrate
rich macromolecules a deep red colour
(magenta).
Acid and Basic dyes PAS will therefore stain up:
glycogen the intracellular storage form of
This table gives some examples of basic and carbohydrate in cells
acidic dyes used in staining. Mucus in cells and tissues, Basement
For basic dyes, the reaction of the anionic membranes, and Brush borders of kidney
groups of cells (these include the phosphate tubules and small and large intestines
groups of nucleic acids, sulphate groups of Reticular fibres (i.e. collagen) in connective
glycosoaminoglycans, and carboxyl groups tissue and Cartilage.
of proteins) depends on the pH at which they In the example shown above, The mucin
are used. produced by goblet cells is stained a purple
For acidic dyes, the dye in question can colour by this stain.
often in addition be selective for particular
acidophilic components. I.e. a technique Masson's trichrome.
This is often used to stain connective tissue.
Tri-chrome - means the technique produces
three colours. Nuclei and other basophilic
(basic-liking) structures are stained blue,
cytoplasm, muscle, erythrocytes and keratin
are stained bright-red. Collagen is stained
green or blue, depending on which variant of
the technique is used.
Alcian blue
Alcian blue is a mucin stain that stains
certain types of mucin blue. Cartilage is also
stained blue. It can be used with H&E, and
with van Gieson stains.
van Gieson
This stains collagen red, nuclei blue, and
erythrocytes and cytoplasm yellow. It can be
combined with an elastic stain that stins
elastin blue/black. It is often used for blood
vessels and skin.
Reticulin Stain
Stains reticulin fibres blue/black. Used with
H&E
Azan
Nuclei are stained bright red, collagen,
basement membrane and mucin are stained
blue, muscle and red blood cells are stained
orange to red. Good for staining connective
tissue and epithelium.
Giemsa.
Usually used for staining blood and bone-
marrow smears. Nuclei are stained dark-blue
to violet, cytoplasm pale blue, erythrocytes
pale pink.
Toluidine blue.
A basic stain that stains acidic components
various shades of blue. It is usually used for
thin acrylic or epoxy sections.
Silver and gold methods.
Sometimes used to demonstrate fine
structures such as cell processes in neurones.
Produces a black, brown or golden stain.
Chrome
alum/haemotoxylin.
Rarely used - stains nuclei blue, and
cytoplasm red. For the pancreas, glucagon
secreting cells are stained pink and insulin
secreting cells are stained blue.
Isamin blue/eosin
Like H&E, but blue is more intense.
Nissl and methylene blue
A basic dye used to stain the rough ER in
neurones.
Sudan Black and osmium.
These dyes stain lipid-containing structures
such as myelin a brownish-black colour.
Immunohistochemical
techniques.
This a specific type of stain, in which primary
antibodies are used that specifically label a
protein, and then a fluoresently labelled
secondary antibody is used to bind to the
primary antibody, to show up where the first
(primary) antibody has bound. A light
microscope, equipped with fluorescence, is used
to visualise the staining. The fluorescent
antibodies are excited at one wavelength of
light, and they then emit light at a different
wavelength. Using the right combination of
filters, the staining pattern produced by the
emitted fluorescent light can be observed. For
example, this photo shows some cells that have
been immunofluorescently stained for the
protein actin.