Kubke Lab:Protocols/Hematoxylin Eosin

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