Technical problems

From:"Pasquetto, Silvia" (by way of Histonet)

Dear all,
in the last days there were problems with computer network in our company
and so I would forward you a message that I have wrote three days ago.
Forgive me if you have received this post twice...

Regards
Silvia Pasquetto
GSK Company

PS. Today I have stained the slides processed with a shorter program (please
find enclose details below) but I find some "interesting" artefacts
consisting in perfect rounded extracellular vacuolization (I hope this word
exist!) of hepatocytes around the centrolobular vessels.

New processation lenghts
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% 45 minutes (instead of 1 hour) at 35C with p/v cycle
5) ethanol 100% 45 minutes (instead of 1 hour) at 35C with p/v cycle
6) ethanol 100% 45 minutes (instead of 1 hour) at 35C with p/v cycle
7) ethanol 100% 1 hour at 35C with p/v cycle
8) toluene 45 minutes (instead of 1 hour) at 35C with p/v cycle
9) toluene 50 minutes (instead of 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 45 minutes (instead of 1 hour) at 58C with p/v cycle
14) wax 1 hour at 58C with p/v cycle


Any suggestions or comments are greatly appreciated

-----Original Message-----
From:	Pasquetto, Silvia
Sent:	marted 18 settembre 2001 17:28
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 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



Silvia Pasquetto
Safety Assessment dept.
GLAXOSMITHKLINE
Tel. +39 +45 921 8111
Int. 4735
sip95711@gsk.com


Silvia Pasquetto
Safety Assessment dept.
GLAXOSMITHKLINE
Tel. +39 +45 921 8111
Int. 4735
sip95711@gsk.com






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