[Histonet] Collagen immunohistochemistry. Enzymatic or heat treatment of frozen sections.

[Guillermo Palao]

  Hello all,
   
  I am working with frozen sections to quantify collagen staining. I am using Anti-Collagen-I Ab from Calbiochem, and they recommend fixing sections after heating or enzymatic treatment with hylaronidase. I have a stock of already acetone-fixed frozen sections where I have tried staining with Anticollagen without success. Before staining I heated fixed sections for 10 min at 42 ºC. I now plan to try hyaluronidase treatment, but I would like to know if anyone has experience staining sections with anti-Collagen once they are already fixed with acetone without previous enzymatic or heat treatment. What is the best way to do it? Is it necessary to cut new tissue and treat it with heat or hyaluronidase before fixation?
   
  As always thank to all you in advance,
   
  Guillermo

		
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