Question

Explain in detail the process of function of staining with Eosin and Hematoxylin? Please use 10-20 sentences

Answer 1

Hematoxylin and Eosin (H & E) staining is the most common staining technique, and it is used for demonstration of nucleus and cytoplasmic inclusions in clinical specimens. Hematoxylin containing alum stains the nucleus light blue. The counterstaining is performed by using eosin which stains the cytoplasm pink color.

Procedure:

Deparaffinize the section : flame the slide on burner and then place it in the xylene. Repeat the treatment.

Hydration : Hydrate the tissue section by passing it through decreasing concentration of alcohol and water bath. (100%, 90%, 80%, 70%)

Stain the sample with hematoxylin for 3-5 minutes.

Then wash the sample in running tap water until sections “blue” for 5 minutes or less.

Differentiate in 1% acid alcohol (1% HCl in 70% alcohol) for 5 minutes.

After that, wash the sample in running tap water until the sections are again turn to blue by dipping in an alkaline solution (eg. ammonia water) followed by tap water wash.

Then stain the sample in 1% Eosin Y for 10 minutes.

After that, wash the sample in tap water for 1-5 minutes.

Then dehydrate in increasing concentration of alcohols and clear in xylene.

Mount the sample in mounting media.

Finally, observe the sample under microscope.