Re: [Histonet] Mouse Testes Fixation and Processing
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.
At 11:18 AM 29/01/2007 -0500, firstname.lastname@example.org 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)
>303 E. Superior Avenue
>Chicago, IL 60611
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