Hematoxylin and Eosin Staining (H and E) Staining

Deparaffinize the section by warming the slides for 5–10 minutes on a slide warming table or by lighting the slides on a burner and then putting them in the Xylene for 3–4 minutes. Repeat the xylene treatment about two or three times with agitation.

Rehydrate the section by passing it through a series of progressively weaker alcohols, starting with absolute alcohol and going down to 30% ethanol. Place the section for 1-2 minutes in each of these alcohol solutions.  Use the tap water to wash, and the distilled water to rinse. Before using dyes to colour the slides, they should be well drained.

Dip the slides in the Coplin jar with the Hematoxylin stain or put the stain on the tissue sections on the slides for 3–5 minutes. Run the slides under running water to clean them.

After washing the slides, put them in the 0.5% (v/v) Hydrochloric acid and take them out (HCl). The nuclei should look dark purple or bluish-red, and the rest of the cell should be white. Rinse the slides for a minute or two in tap water.

Now, dip the slides more than once in the ammonia water. The part of the section should be blue. Rinse the slides in tap water.

Dehydrate the sample by going from 50% Ethanol to 70% Ethanol to 90% Ethanol to 95% Ethanol in ascending order. Put the sections in each of these alcohol solutions for 30 seconds to 1 minute, and then rinse it with Absolute Ethanol.

Dip the slides for 30 seconds to 1 minute in the Eosin solution. Absolute alcohol should be used to rinse the slides.  Place the slides two times in Xylene solution for 30 seconds to 1 minute each. Drain off any extra Xylene, attach the DPX, and put the coverslip on top. Let it dry, then look at it with a microscope.