STAINING

Staining

widely used in histopathology and diagnosis, as it allows for the

identification of abnormalities in cell count and structure under the microscope

metachromasia

when certain stains change the coloration of cells and tissues significantly, different from the colour of the original dye complex

paraffin

which section of tissue are normally used for staining?

Xylene

makes the tissue rehydrated and then made translucent using a clear substance called what? before staining

Hematoxylin and Eosin

the most widely used staning technique in histopathology

Hematoxylin and Eosin

This is the most common histologic stain, used to differentiate different tissue structures

Hematein

In the H&E stain, a mixture of oxidised hematoxylin known as ____ is used.

salts of aluminium, iron and tungsten

most commonly used mordants

Hemalum

most commonly used mordants and this substance is known as what

Nuclei blue

Hemalum stains what colour

Eosin

stains proteins and cytoplasm varying shades of pink

chemical attraction between tissue and dye

The principle behind H & E stain

Hematoxylin

a basic dye imparts blue-purple contrast on basophilic

structures, primarily those containing nucleic acid moeties such as

chromtatin, ribosomes and cytoplasmic regions rich in RNA.

Eosin

counterstains the basic elements such as RBCs, cytoplasm, muscle and collagen in varying intensities of pink, orange and red

1 gm hematoxylin in 10 ml ethanol

10 gm ammonium alum in hot distilled water

mix and boil and add 0.5 gm of mercuric oxide and filter

requirements for Harris hematoxylin stain

Yellow eosin 1 gm

Distilled water 80 ml

Ethanol 320 ml

Glacial acetic acid 2 drops

requirement for Eosin solution

Deparaffinization

flame the slide on burner and place in the xylene. Repeat the treatment

to remove the wax.

Hydration

Drain xylene and hydrate the tissue section by passing through decreasing

concentration of alcohol baths (100%, 90%, 80%, 70%) and water.

Nuclear Staining

Stain in hematoxylin for 3-5 minutes. then each for 5 or less minutes

Differentiation

selective removal of excess dye from the section). Dip in 1% acid alcohol

(1% HC1 in 70% alcohol) for a few seconds.

Blueing

Rinse i n running tap water. Dip in ammonia water until the sections become blue, followed by tap water wash.

Counterstain

Stain in 1% Eosin Y for 10 minutes then wash for 1 to 5 minutes

Dehydration

Dehydrate in increasing concentration of alcohols

Clearing

Put slides in two xylene baths for clearing

Mounting

Mount in DPX or other mounting media

Blue, black

colour of nuclei

Pink/purplish pink

colour of cytoplasm

Deep red

colour of muscle fibres

Orange red

colour of RBC

Dark blue

colour of calcium

Grey blue

colour of mucin