[Histonet] Silly question about deparaffinization

[Rittman, Barry R]

Eva
I would suggest that you do a trial and error run.
Deparaffinization time with xylene will depend on several factors
including: thickness of sections, type of paraffin, number of slides,
agitation, temperature of solution, volume of fluid, number of times
that xylene bath was used, quality of the xylenes solution and so on.  
I have always erred on the conservative side and given a longer time as
you have done in the past. However, for many labs, as Linda has
indicated, 3 minutes in each bath is fine. 
Suggest that you try a set of slides and deparaffinize them for 3, 5 7
and 10 minutes and compare your results.
Barry

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sebree
Linda A.
Sent: Wednesday, June 29, 2005 9:14 AM
To: Eva C Andersson; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Silly question about deparaffinization

We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95%
ETOH and 1 70% ETOH, all for 3 minutes each.  We never skimp on the
xylene times but have been known to shorten the ETOH times if we
manually agitate the slides and are in a real hurry.

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
DM223-VA
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX:  (608)262-7174



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eva C
Andersson
Sent: Wednesday, June 29, 2005 8:57 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Silly question about deparaffinization


Good morning,
I am probably asking a very silly question but I recently spoke to 
someone about deparaffinization and hydration. I found that the times 
that they use and the times that we use are very different. So my 
question to you is this: What lenghts of time do you use for Xylenes, 
100% alcohol, 95% alcohol and 70% alcohol?
It is about deparaffinising charged (+) slides. So nothing fancy. Just
curious. Maybe I can cut back some on the timed that we use which are:
30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your
responses. Eva Andersson Georgetown University

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