[Histonet] get back eosin stain?

[Yak-Nam Wang]

Hello,

I have a colleague with a staining problem. He has cells grown on polymeric 
scaffolds. He can not embedd in paraffin due to a mismatch in the processing 
and his scaffold material; he has thus embedded in OCT.

This is his procedure:
Fix cell seeded scaffold in formalin (these are delicate samples thus, fixing before embedding)
Cryoprotect in grade sucrose to 30% sucrose
Embed in OCT
Section at 6um
General H&E staining-using 1 min for Hematox and 1.5 min for Eosin (following 
one of Richard Allan suggested protocols).

Problem: there is no eosin staining. The cells are just purple, there is no pink at all. We are not sure why this is. Is this caused by fixation? 

Does someone have any suggestions on how he can get the classic H&E stain look for these samples? He has tried fresh solutions as well as running other tissue (unfixed) through. It doesn't seem to be the solutions used.

Any suggestions would be appreciated. Thank you
Yak-Nam


Reserach Associate
CIMU
University of Washington
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