[Histonet] Mouse Testes Fixation and Processing

[Laurie Reilly]

Donna and All,
Bouin's is the fixative of choice for testis, and my experience with 
formalin fixation of testis tells me that your observations are correct.
A formalin-acetic acid-ethanol mix may be better. Maybe Davidson's fixative?

Another "trick of the trade" is to start the processing of testis (and 
other very delicate tissues) in 30% Ethanol then 50%, 70%, 80%, 90%, 95%, 
100%, 100% then 50:50 EtOH:Xylene, 2x Xylene 3x Paraffin.

For mouse tissues 15 minutes in each of these reagents would be enough.

Regards,   Laurie.


At 11:18 AM 29/01/2007 -0500, djemge <@t> aol.com wrote:
>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
>312-503-2036
>d-emge <@t> northwestern.edu
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Mr.Laurie Reilly
School of Veterinary and Biomedical Sciences
James Cook University
Townsville. 4811
Australia.

Phone  07 4781 4468
Fax      07 4779 1526