Hi There,
Is anyone using this technique to stain chlamydia in placental samples
(working with sheep here)? I was looking for a working technique but
it looks like it is not easy to find.
Thanks for your thoughts!
Julio
Rene J Buesa via Histonet <histonet@lists.utsouthwestern.edu> escribió:
Hi Mike:The steps you desrcibe are wrong.After you finish staining
your PAP smear, just wash them in the last ethanol → oven dry at
60ºC for 5 minutes or as required if the smear is too thick and when
completely dried → coverslip.This final drying has to take place at
temperatures above room temp. because you have to be absolutely sure
the smear (or stained tissue section) is absolutely dried before
applying the mounting medium → coverslip.The steps you write about
2-propanol and mineral oil are those the tissues have to go through
before paraffin infiltration. René
On Tuesday, May 24, 2016 6:36 PM, Mike Toole via Histonet
<histonet@lists.utsouthwestern.edu> wrote:
René and other Histonetters,
After reading the paper:
Buesa RJ, Peshkov, MV. Histology without xylene. Ann Diagn Pathol.
2009 Aug;13(4):246-56. Epub 2009 Feb 5.
It appears that xylene in the final clearing steps is replaced with
isopropanol and mineral oil as follows:
· 5:1 isopropanol to mineral oil 50°C
· 2:1 isopropanol to mineral oil 50°C
· Undiluted mineral oil 50°C
· Drying oven 5 minutes 60°C
· Coverslip
Do you feel these clearing steps be applied to the pap stain in
order to eliminate xylene? If so, can it be done at room temperature?
Thanks,
Mike
Mike Toole, BS, CT(ASCP)CM
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