Hematoxylin and Eosin Staining Procedure Steps

Prepare Slide

Deparaffination

Xylene (3 changes for 5 minutes each)

Rehydration

Hydrate sections through graded alcohol

Wash in deionized water for 1 minute

Nuclear stain (Hematoxylin 15 minutes)

Tap water

Differentiation (1% acid alcohol; 1-3 rapid dips)

Tap water

Bluing (Ammonia water; 10-15 dips)

Running water

Counterstain (Eosin)

Dehydration (95% Ethyl Alcohol; 15-20 dips)

Dehydration (95% Ethyl Alcohol; 15-20 dips)

Dehydration (95% Ethyl Alcohol; 15-20 dips)

Dehydration (95% Ethyl Alcohol; 15-20 drops)

Clearing (Xylene; 15-20 dips)

Clearing (Xylene; 15-20 dips)

Clearing (Xylene; 15-20 dips)

Air dry

Mount