H-E stain

 

Intended use:

It is mainly intended to display the common morphological structure for normal components of various tissues and pathological changes. H-E stain is the basic and necessary method in biology, histology, pathology and cytology. It is widely used in diagnostics, teaching and research, with important values.

 

Principle:

The nucleolus, which is made up of acid materials, has a strong affinity for alkaline stain (Hematoxylin) while the cytoplasm, made up of alkaline materials, binds favorably to acid stain (Eosin). Therefore, in the presence of the H-E stain, nucleolus will be stained bright indigo blue by hematoxylin; cytoplasm, stroma, muscle fiber and collagen fiber will be varying shades of pink; and erythrocyte will be salmon pink.

 

Methods:

1. Deparaffinize tissue in two washes of xylene for 5 minutes each. Wash tissue twice in 95% ethanol for 1 minute each.

2. Place tissue in 80% ethanol for 1 minute, then rinse with tap water for 1 minute.

3. Stain for 3~5 minutes with Harris. Rinse with tap water for 1~2 minutes.

4. Place tissue in 0.5%~1% hydrochloric acid ethanol for a few seconds.

5. Wash the 'Blue' tissue in tap water, or with lithium carbonate solution, for 5~10 minutes, followed by another water rinse for 1~2 minutes.

6. Place tissue in Eosin for 30~60 seconds. Rinse with water.

7. Dehydrate tissue in 80% and 95% ethanol, respectively.

8. Dehydrate in 100% ethanol for 1~2 minutes, then clear in phenol xylene and xylene for 1~2 minutes each.

9. Mount with mounting media and examine microscopically.

 

Specifications:

 

Contents	2Btlsx100ml/kit	2Btlsx1000ml/kit	Components
Harris Hematoxylin Solution	1x100ml	1x1000ml	Hematoxylin
Eosin stain	1x100ml	1x1000ml	Eosin
Hematoxylin&Eosin  stain (Mix)	4BTx250ml  (Hematoxylin and Eosin are 2 BT respectively)		Hematoxylin, Eosin

 

Precaution:

1. It is normal if a layer of oxidative film is present on the surface of Harris Hematoxylin and some crystal precipitates of aluminum sulphate at the bottom of Harris. Prior to use, however, the oxidative film must be removed by filtration. Filter regularly.

2. If staining with Hematoxylin at lower room temperature becomes difficult, extend the staining time as needed.

3. Differentiation refers to the washing off of excess hematoxylin adsorbed by cells with hydrochloric acid for a brighter contrast between karyon and cytoplasm. It cannot last too long, or else the karyon stain will be weakened.

4. After staining with Eosin, do not overwash the smears for too long. Otherwise, Eosin discoloring may occur. Use low-concentration ethanol for best results.

5. The H-E staining for printing slice and smears, after fixation and water wash, should follow the same procedure as that of paraffin tissue from step 3.

6. Good H-E stain tissue depends much on the timely and sufficient fixation.

7. Do not use the reagents beyond the stated expiration date. For kit storage, avoid exposure to extreme high or low temperature and sunlight.

 

Expected Results:

Nuclei appear indigo blue while cytoplasm, stroma and fibres appear varying shades of pink.