[Histonet] Cryosectioning, fixing and H&E staining

From:chau0056

 
Hello,
     I want to cryosection collagen (bovine type I) matrixes seeded with
human stromal fibroblasts.  I am looking at collagen production by the
fibroblast.  My two questions are 1) Is it better to fix before or after
cryosectioning?  I’m using a 1% glutaraldehyde- 10% formalin fixative.  2)
After section I want to use a Hematoxylin (Modified Harris) and Eosin Y
alcoholic stain, but I’m having trouble seeing the Eosin stain.  I have
tried shortening the destaining step, but it doesn’t seem to be working. 
Also, the staining steps, some of my samples don’t want to stay on the
slide.  

Eric
Department of Mechanical Engineering
University of Minnesota       



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