[Histonet] RE: reduction/elimination of red cell autofluorescence

C.M. van der Loos c.m.vanderloos <@t> amc.uva.nl
Tue Aug 25 02:30:19 CDT 2009

Date: Mon, 24 Aug 2009 10:58:43 -0400
From: "Thurby, Christina" <christina.thurby <@t> bms.com>
Subject: [Histonet] reduction/elimination of red cell autofluorescence
on FFPE sections for IF examination
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>

Can anyone give feedback on reagents/procedures to use for immunofluorescence to reduce/eliminate red cell autofluorescence on FFPE sections.  I am using an indirect method of labeling for two antibodies (FITC and Texas Red).

I have not tried 0.1% sodium borohydride.  Will this help for formalin fixed specimens?  I have read that it is used for gluteraldehyde autofluorescence reduction.  If appropriate how long should this reagent be applied to the specimen and at what temperature.
Christina Thurby
christina.thurby <@t> bms.com
Dear Christina,One of the most spectacular ways of getting rid of autofluorescence in FFPE tissue sections is 'unmixing' the autofluorescence from real fluorescence. This can be done with spectral imaging using the Nuance system (http://www.cri-inc.com/applications/fluorescence.asp). Obviously this is not affecting your epitopes with all kinds of nasty chemicals. On top of that, using the Nuance system, the autofluorescence can be pseudo-stained in grey as a kind of 'counterstain'. It's absolutely worth looking at it! Spectral imaging is the subject of workshop #66 at the upcoming NSH convention.Cheers, Chris Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone:  +31 20 5665631
fax:    +31 20 6960389
e-mail: c.m.vanderloos <@t> amc.uva.nl

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