RE: Technical problems

From:"Philopena, Jennifer"

Sylvia,
When I began processing mouse and liver tissue, I started with a processing
protocol similar to yours - an hour in each reagent.  My rodent tissue came
out cracked and brittle.  I decreased my processing schedule to 20 minutes
per reagent.  My tissues look a lot better.  I also make sure to gross the
tissue at 4-5mm thickness.
Jen




-----Original Message-----
From: Pasquetto, Silvia [mailto:sip95711@GlaxoWellcome.co.uk]
Sent: Thursday, September 20, 2001 12:57 PM
To: HistoNet@pathology.swmed.edu
Subject: Technical problems


Hi guys!
I need all you for some technical problems regarding liver slides
preparation. I work with animal (rat) tissue.
Recently, I have processed some pathologic (i.e. oil red o' staining
revealed fatty accumulation) rat liver samples and I was amazed when after
processing, cutting and staining of these samples there were some cracks in
tissue under microscopical observation. The appearance is similar to the
fissures in the dry land: I hope that comparison is clear for you! I think
there is a correlation between this appearance and the length of
processation of these sensitive livers. What do you think about this?
Tomorrow I have tried to reduced the lenght of  ethanol absolute, toluene
and wax staying. Please find below the details of the first processation
that gave me this problem.

1) formalin
2) ethanol 70% 1 hour at 35C with p/v cycle
3) ethanol 95% 1 hour at 35C with p/v cycle
4) ethanol 100% 1 hour at 35C with p/v cycle
5) ethanol 100% 1 hour at 35C with p/v cycle
6) ethanol 100% 1 hour at 35C with p/v cycle
7) ethanol 100% 1 hour at 35C with p/v cycle
8) toluene 1 hour at 35C with p/v cycle
9) toluene 1 hour at 35C with p/v cycle
10) toluene 1 hour at 35C with p/v cycle
11) wax 45 minutes at 58C with p/v cycle
12) wax 45 minutes at 58C with p/v cycle
13) wax 1 hour at 58C with p/v cycle
14) wax 1 hour at 58C with p/v cycle

In your opinion which is the correlation between this troublesome effect and
the fatty accumulation in the liver?
Any suggestions or comments are greatly appreciated

Thanks in advance
Silvia Pasquetto




***************************************************************
 This message and any attachments is solely for the intended recipient. If
you are not the intended recipient, disclosure, copying, use, or
distribution of the information included in this message is prohibited --
please immediately and permanently delete this message.




<< Previous Message | Next Message >>