[Histonet] Ideal fixation of mouse prostate

[Rene J Buesa]

I don't think that your tissues are overfixed (under separate cover I am sending an article on the subject of fixation time) but I think it is a very bad idea to leave the tissues overnight in alcohol (even worse at 25%).
Once the tissue as fixed (as you do now) the processing should start and the dehydration should be shorter than for human tissues because mouse tissues have less water than human and will tend to get very friable and dificult to cut.
Also xylene is a proble. I will send you a protocol with mineral oil under separate cover.
René J.

--- On Thu, 2/26/09, Vanessa J. Phelan <vjp2105 <@t> columbia.edu> wrote:

From: Vanessa J. Phelan <vjp2105 <@t> columbia.edu>
Subject: [Histonet] Ideal fixation of mouse prostate
To: histonet <@t> lists.utsouthwestern.edu
Date: Thursday, February 26, 2009, 3:40 PM

Hi everyone,

I am very new to histonet, actually just joined.I hope you guys can help. I
was wondering if anyone has any experience in the fixing and processing of
mouse issues, namely prostate or bladder. I am currently trying to optomize
protocols for this as I have very recently taken on the position of
histology manager in a research lab, before this I worked with human
samples.
We have the VIP processor, and the protocol being followed at the mo is,
fixation overnight (though the tissues are tiny) and then washed in PBS and
then into 25% alcohol (sometimes over night) and then into the processor,
the cycle starts with alcohol instead of formalin.
I fear that the tissues are being overfixed and also does anyone leave their
tissues in 25% alcohol before processing?

Thanking you in advance for any input,

Nessa


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