[Histonet] Re: Autofluorescence reagent

[ALAN K MEEKER]

From: "Andrea T. Hooper" <anh2006 <@t> med.cornell.edu>
===
>>At 11:37 AM -0600 8/18/05, Gayle Callis wrote:
>I think the Chemicon autofluorescence removal reagent - whatever 
>they call it, is to get rid of lipofuscins - their website will have 
>the information how it works.

Has anyone used this? Have you used this yourself Gayle? It worries 
me that the instructions say to use it after staining and then 
subsequently do 70% ethanol rinses!
===
Andrea, I have tried the Chemicon product a couple of times. We were looking for a way to eliminate autofluorescence in probing DNA sequence using a Cy3-labelled PNA probe on various FFPE tissues. We followed the manufacturer's directions and saw a reduction in background autofluorescence to varying extents (depending on the tissue type). I'd say at best we might have lost about 1/3 of the background. Unfortunately, this was accompanied by a decrease in the fluorescent intensity of the Cy3 probe signals as well, so we really dodn't gain anything. I didn't do any controls without the reagent to see if this reduced signal was due to the reagent, of to the ethanol steps which we usually do not do in our protocol.


Alan Meeker, PhD
Assistant Professor of Pathology
Department of Pathology
Division of Genitourinary Pathology
Bunting-Blaustein Cancer Research Building  Room 153
1650 Orleans Street
Baltimore, MD 21231-1000

ph (410) 502-7880 /(410) 614-5686
fax (410) 502-9817