Re: Fixation and Processing

From:louise renton

Dear Travis,
To the first part of your question:
If the specimens were properly fixed the first time around, there is no need 
to hold them in formalin. The mushiness is due to poor dehydration causing 
ultimately poor (nearly impossible) infiltration by wax. Therfore, I would 
suggest that you reverse process to  80% or 95% alcohol and start again from 
there. It is a terrible pain to do isn't it :>)!

I rely on someone with more immuno experience to answer you on the 2nd part 
of your query. My gut feeling is that keeping it in 70% shouldn't be too 
bad, but I'm interested in what others will say.

Best regards
Louise Renton
Bone Research Unit
South Africa
Tel & fax +27 11 717 2298
"Time flies like an arrow, fruit flies like a banana"

>From: Peterson Lab 
>Subject: Fixation and Processing
>Date: Tue, 22 Oct 2002 10:06:22 -0500
>My question today is as follows.  Our tissue processor was set for weekend 
>run last night and when I came in at 4:30, the processor hadn't started 
>yet.  I ran a short run on the processor consisting of 10 minutes in each 
>step in order to process the tissue for the day.  Many of the bigger pieces 
>of tissue didn't cut well at all and were mushy.   I plan to reprocess 
>these tissues and was wondering how long to leave them in formalin when our 
>processor run for today doesn't start until 6:00 tonight.  My other 
>question is that one of the tissues was a breast tissue that was positive 
>and needs immunostains.  Do I put these tissues in 70% ethanol all day and 
>then put them on the processor?
>I know that this is a long question, but I thought it necessary to explain 
>the situation in detail.  Thanks so much for your time!
>Travis Troyer

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