[Histonet] Silly question about deparaffinization

[Pamela Marcum]

Eva,

Depending on the thickness of the sections my times can vary. Sections of 4 
to 6 micron are deparaffinized in our laboratory at 2 changes at 5 minutes 
each with alcohols at 3 minutes each per station to water.  Sections 
thicker than 6 microns may go as long 15 minutes X 2 in xylenes and 5 to 10 
minutes in each alcohol.  I am very paranoid about getting all of the 
paraffin (or MMA where I use one hour per change for xylenes) so I may go 
over board.  I also change my reagents more often in humid weather.  I do 
not use an automated stainer so agitation only takes place at the beginning 
of the changes to assure the last reagent is rinsed off.

Hope this helps.

At 09:56 AM 6/29/2005, Eva C Andersson wrote:
>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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Best Regards,

Pamela A Marcum
Manager, Histology Special Procedures
University of Pennsylvania
School of Veterinary Medicine
R.S. Reynolds Jr.  CORL
New Bolton Center
382 West Street Road
Kennett Square, PA 19348

Phone - 610-925-6278
Fax     - 610-925-8120
E-mail - pmarcum <@t> vet.upenn.edu