[Histonet] AFB Fites of cytospins

[Rene J Buesa]

Carrie:
The cytospin will not need dewaxing, as you point out, but I do not foresee any problem when compared with the control. You have to realize that dewaxing the control is intended only to expose the tissue section to the reagents.
As to the peanut oil if you are trying to stain for "common" Mycobacterium it is not necessary because their cover is quite strong. If you are trying to detect M. leprae it is when you use the mixture of peanut oil (or any other vegetable oil for that matter) mixed with xylene. The rationale is that the oil will "protect" the delicate cover of the organism.
But again, not even for M. leprae you will need the mixture of oil and xylene if you are using a cytospin, because they do not require "deparafinization" (dewaxing).
René J.

--- On Mon, 5/18/09, Carrie Disbrow <disbrc <@t> shands.ufl.edu> wrote:


From: Carrie Disbrow <disbrc <@t> shands.ufl.edu>
Subject: [Histonet] AFB Fites of cytospins
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, May 18, 2009, 6:12 PM


Hello Histonet.
Has anyone had experience and good results doing AFB Fites on cytospin slides from BAL's?
Do you use the peanut oil/xylene (PO/Z) solution or just 100% peanut oil? Or does the cytospin slide need any peanut oil?
After the deparaffinization step, if it can be called that, we are going to stain using a Dako Artisan special stainer. One problem I foresee is that the control will be de-arffinized in the traditional PO/Z while the patient slide may not.
Also, I was told that Cytolyte contains alcohol and asked that the slides not be prepared using Cytolyte.
Thanks for your comments. Please respond back to the histonet if possible and not my email address. 
Carrie
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