Re: [Histonet] Ovary fixation

From:Rene J Buesa

Try infiltration with a mixture of ethanol (E) + isopropanol (I) + mineral oil (M) at:
  E:I:M (2:3:1) at 45C for 1.50 hours followed by E:I:M (1:3:2) at 50C for 1.25h
  Complete with 4 paraffin baths at 58C of 0.75, 0.30, 0.30 and 0.50 h
  No xylene or substitutes are needed. More gentler and better infiltration is obtained.
  Ren J.

Heidi Miers  wrote:
  I section dog and cat ovaries on a regular basis and am unhappy with the 
quality of the follicles. The primordial follicles are found near the surface 
and when I look at them under the microscope they all look like they have 
vacuoles in the oocytes. Some ovaries come out looking OK. I am wondering if 
anyone has suggestions. Could it be fixation? We use 10% formalin for at least 
3 days. How about processing? Could it be that the xylene substitute I use 
isn't clearing to allow proper paraffin infiltration?
Also, for those who use a regressive H&E-what type of eosin and for how long 
to you let it stain?
Thanks, Heidi

Research Specialist, Sr.
Imaging and Histology Core Facility
Northern Arizona University

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