[Histonet] Mouse Testes Fixation and Processing


Hello All. I am having an ongoing problem with formalin fixed, paraffin embedded whole mouse testes (Adult mouse and Pups). I need to get this issue resolved and have consistent results for my investigators. They are using these testis for xgal, IF, IHC and In Situ. All my Bouin's fixed testes looks beautiful but the formalin fixed testes are horrible. Bouins is not suitable for much of what they are doing. I have tried several processing schedules and nothing seems to work. Sometimes the FFPE's seem very dry and shrunk, sometimes the cells look like they have big halos around them and you can't distinguish them enough for staging. This is frustrating! All other tissue seems to be fine except these and there is a big variability in how these testes turn out. They are NBF or cold 4%PFA PBS fixed for 24 hours. Typical processing schedule is either 30 minutes each station or 1 hour each station (formalin, 2x70%, 80%, 2x95%, 2x100% reagent alcohol, 2x Xylene, 3x Ameraffin Paraffin (Cardinal cat# M7346-1A). 
Donna J. Emge, HT(ASCP)
Northwestern University
Lurie 7-220
303 E. Superior Avenue
Chicago, IL  60611
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