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