Re: [Histonet] FISH cytology fixatives

In Cytological preparation, one would use an acetic acid in the fixative to lyse the RBC's, which could ultimaetly interfere with the diagnosis.? Journal articles are great to help you research and give you a starting point for what it is you want to do, that is where I started.? But when you start doing the procedure, or playing with until you get it to work, you always need to start with the suggestions of the manufacturer of the probe.? That is how they validated it and it is known to work under those conditions.? At that point you can play with fixatives, if you need to.

A lot of FISH testing requires post fixation anyway.? For, instance, the tissue comes in formalin (or whatever), but after you de-wax the slides you have to begin the pre-treatment, an most of the time there is a step in there that involves placing the slides in formalin (or whatever).


-----Original Message-----
From: Kim Merriam 
To: Histonet 
Sent: Wed, 27 Feb 2008 7:42 am
Subject: [Histonet] FISH cytology fixatives

Hi Everyone,
  I am working up some FISH staining protocols for cytology (cell culture) 
slides (eventually to be OCT and FFPE tissues, but I need to walk before I can 
  Most of the journals that I have read recommend using a fixative containing 
methanol and acetic acid (3:1).  The DAKO probe I am using calls for the use of 
3.7% fomaldehyde.  I am wondering why acetone (or acetone/ethanol) is not the 
routine fixative for such a procedure.  Can someone let me know why to choose 
one fixative over another and what the merits of methanol/glacial acetic acid 
would be?
  Thanks in advance,

Kim Merriam, MA, HT(ASCP)
Cambridge, MA
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