I am staining ovine hard tissue sections of Resin embedded samples with some soft tissue components. The samples are approximately 20 um thick

The hematoxylin is very overpowered and non-specific in the section.

Here is my protocol:

3mins - Acetone etching (removing resin)

3mins - Acetone etching (removing resin)

Hydration:

5mins - 100% Ethanol

5mins - 100% Ethanol

5mins - 90% Ethanol

5mins - 70% Ethanol

5mins - DH2O

1mins - Hematoxylin (modified Myers)

Dehydration:

10mins - DH2O running water

15 secs - 70% Ethanol rinse

5mins - 90% Ethanol

5mins - 100% Ethanol

5mins - 100% Ethanol

10mins - Eosin (Y Solution)

Wash - 100% Ethanol

Wash - 100% Ethanol

5mins - 100% Ethanol

5mins - 100% Xylene

5mins - 100% Xylene

Mount slides

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