RE: tissue processing

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From:"Saby, Joseph" <> (by way of histonet)
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Dear Jennifer-

One thing you need to change for sure:  do not store your brains in 70%
ethanol, as this will cause an artifact in and around certain structures in
the brain.  It has been years since I've seen the article describing this so
I cannot give you chapter and verse, but I can say from experience that I
once had this artifact when processing rodent brains over the weekend.

Your processing times and stations sound right, but again I think storing
your samples in 70% ethanol could be causing your problem here.  I do not
have a suggestion for your right off the top, but I'm sure someone else in
Histonetland will be delighted to help you!


Joseph A. Saby, BA HT(ASCP)
Diagnostic Pathology, Pathology & Experimental Toxicology
Parke-Davis/Int. Pharm. Res. Div. Warner-Lambert
Ann Arbor, MI 48105
Phone: (734) 622-3631
Fax:     (734) 622-5718

-----Original Message-----
From: Mequita Praet []
Sent: Wednesday, November 17, 1999 6:26 PM
To: Philopena, Jennifer
Cc: 'HistoNet Server'
Subject: Re: tissue processing

I have not worked with animal tissue before but I do work with skin
They become brittle if I over dehydrate. I would suggest you should try
eliminating one of your absolute alcohols.

Philopena, Jennifer wrote:

> Hi.  I work with a Shandon Citadel 2000 and I (try to) process rat and
> tissues (liver, lung, spleen, kidney, brain, heart).  The livers and
> that I process are consistently very dry.  This is my procedure: 24 hours
> 10% neutral buffered formalin, store in 70% reagent alcohol, 20min in 80%
> reagent alcohol, 2X 20min in 95% reagent alcohol, 3X 20min in 100% reagent
> alcohol, 3X 20min in Propar (Xylene substitute), 2X 20min in paraffin
> (Paraplast Plus).  I'd like to fix this problem with the tissues being
> over-dehydrated.
> Thanks.
> Jennifer Philopena

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