Kubke Lab:Protocols/Hematoxylin Eosin
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Hematoxylin and Eosin staining
WORK IN PROGRESS --- EXPECT FREQUENT CHANGES
Modified Hematoxylin and Eosin staining procedure for chick embryo tissue
(Provided by Satya Amirapu,procedure not yet optimised)
Method | |
1.Tap water | Approximately 10 dips |
2.Absolute alcohol(ethanol)1 and 2 each | 1 dip each |
3.Tap water | Approximately 10 dips |
4. Haematoxylin Gils2 | 45sec - 1.5 minutes |
5. Differentiate in 1% acid alcohol | 1 Dip |
6. Tap water | Approximately 10 dips |
7. Blue in 1% Lithium carbonate | 6 dips |
8. Tap water | Approximately 10 dips |
9. Stain in 1% Eosin | 10-30 seconds |
10.Absolute alcohol 1 and 2 | Rapid dips |
11.Xylol 1,2 and 3 | 10 dips each |
12.Apply DPX to your slides and coverslip |
Preparing the Solutions
1% ACID ALCOHOL
- 3.5L Absolute alcohol 850ml
- 1.5L distilled water 350ml
- 25ml HCL 6.5ML
Mix together and store in a cool place
Gils 2 Haematoxylin
- Distilled water 730ml
- Ethylene Glycol 250ml
- Pure Haematoxylin 4gms
- Sodium Iodate 0.4gms
- Aluminium sulphate 70.4gms
- Glacial acetic Acid 20ml
1% Eosin
- EosinY- 20gms
- Distilled water- 2L
- 1% Calcium Chloride 20ml
Dissolve Haematoxylin and Alum separately in distilled water and then mix.
1% Lithium Carbonate
- Lithium carbonate 1gm
- Distilled water 100ml
References
Disbrey & Rack (1970), Histological Laboratory Methods, E.&S.Livingstone, Edinburgh and London