H&E Chemistry

The H&E stain uses two dyes, hematoxylin and eosin. This combination is used as the dyes stain different
tissue elements.
Hematoxylin reacts like a basic dye with a purplish blue colour. It stains acidic, or basophilic, structure
including the cell nucleous (which contains DNA and nucleoprotein), and organelles that contain RNA
such as ribosomes and the rough endoplasmic reticulum.
Eosin is an acidic dye that is typically reddish or pink. It stains basic, or acidophilic, structures which
includes the cytoplasm, cell walls, and extracellular fibres.
Dye origins
Hematoxylin is extracted from the logwood tree and purified. It is then oxidized and combined with a
mordant (typically aluminium) to allow it to bind to the cell structures. Of the many hematoxylin
preparations used in histology Gills hematoxylin, Harris's hematoxylin and Mayer's hematoxylin are the
most popular.
Eosin is formed by a reaction between bromine and fluorescein. There are two eosin variants typically
used in histology: eosin Y which is slightly yellowish and eosin B which is slightly bluish. Eosin Y is most
popular.
used in histology: eosin Y which is slightly yellowish and eosin B which is slightly bluish. Eosin Y is most
popular.

Figure 6: Hematoxylin chemical structure

Figure 7: Eosin Y chemical struct

Some Common Special Stains
The images below illustrate some of the common special stains and their applications.

Figure 8: Masson's Trichrome (skin). This stain is intended for use in histological
observation of collagenous connective tissue fibers in tissue specimens. It is used to assist
in differentiating collagen and smooth muscle in tumors and assists in the detection of
diseases or changes in connective/muscle tissue.

Figure 9: Modified GMS Silver Stain (Left: Pneumocystis, lung) (Right: Aspergillus
infection, lung). The Modified GMS Silver stain is intended for use in histological
observation of fungi, basement membrane and some opportunistic organisms such as
pneumocystis carinii in tissue specimens.

.

Figure 10: Periodic Acid Schiff (kidney). PAS staining is mainly used for staining
structures containing a high proportion of carbohydrates such as glycogen,glycoproteins,
proteoglycans typically found in connective tissues, mucus and basement membranes.
Often used to stain kidney biopsies, liver biopsies, certain glycogen storage diseases in
striated muscles and suspected fungal infections.

Figure 11: Perls Prussian Blue Iron (liver). This stain is used to detect and identify ferric
(Fe3+) iron in tissue preparations, blood smears,or bone marrow smears. Minute amounts
of ferric iron (haemosiderin) are commonly found in bone marrow and in the spleen.
Abnormal amounts of iron can indicate hemochromatosis and hemosiderosis.
.

Figure 12: Ziehl Neelsen (Acid Fast Bacillus, lung). This stain is used to detect and
identify acid fast bacilli in tissue. Bacilli are rod-shaped bacterial organisms. A primary
function of this stain is to identify tuberculosis in lung tissue.

Figure 13: Alcian Blue (intestine). Alcian Blue is normally prepared at pH 2.5 and is used
to identify acid mucopolysaccharides and acidic mucins. Excessive amounts of non-
sulfated acidic mucosubstances are seen in mesotheliomas, certain amounts occur
normally in blood vessel walls but increase in early lesions of atherosclerosis.




Figure 14: Alcian Blue and PAS (intestine). A stain that combines the properties of both
Alcian Blue and Periodic Acid Schiff staining.

Figure 15: Gomori Trichrome (blue) (submucosa). Trichrome stains are used to stain and
identify muscle fibers, collagen and nuclei. They can be used to contrast skeletal, cardiac
or smooth muscle. The Gomori Trichrome is a simplification of the more elaborate
Masson trichrome stain and combine the plasma stain (chromotrope 2R) and connective
tissue stain to provide a brilliant contrasting picture.




Figure 16: Gomori Trichrome (green) (submucosa). Trichrome stains are used to stain
and identify muscle fibers, collagen and nuclei. They can be used to contrast skeletal
,cardiac or smooth muscle. The Gomori Trichrome is a simplification of the more
elaborate Masson stain and combines the plasma stain (chromotrope 2R) with the
connective tissue stain to provide a brilliant contrasting picture.