Hematoxylin and Eosin Stain for Glycol Methacrylate Sections

Technovit Glycol Methacrylate: Technical Information and Help From Heraeus Kulzer

Hematoxylin and Eosin stain is a good general stain for many types of tissue.

Solutions:

Gill's Hematoxylin

• Hematoxylin (C.I. 75290) - 6 g
• Sodium Iodate - 0.6 g
• Aluminum Sulfate - 52.8 g
• Distilled Water - 690 ml
• Ethylene Glycol - 250 ml
• Glacial Acetic Acid - 60 ml

Eosin

• Eosin Y (alcoholic C.I. 45380)- 0.5 g
• Ethanol 96% - 100 ml
• Glacial Acetic Acid - 2 drops

Acid Ethanol

• 4 ml of 25% HCl.
• 100 ml of 70% Ethanol.

Procedure:

1. Stain in Gill's hematoxylin for 15 minutes.
2. Wash in tap water for 10 minutes.
3. Decolorize plastic, if necessary, with acid alcohol followed by tap water rinse.
4. Rinse in distilled water.
5. Scott's Tap Water for 1 - 2 minutes.
6. Two changes of distilled water.
7. Counterstain with Eosin for 2 - 5 minutes.
8. Dehydrate using 96% and 100% ethanol.
9. Clear in xylene and coverslip.

Results:

Nuclei - blue
Basophilic cytoplasm - blue
Acidophilic cytoplasm - pink
Muscle tissue - pink
Connective tissue - pink

Citation:

Gerrits, P. O. and Smid, L., "Staining Procedures for Tissues Embedded in 2-Hydroxyethyl Methacrylate", Heraeus Kulzer.