Date: Mon, 24 Aug 2009 10:58:43 -0400 From: "Thurby, Christina" <christina.thu...@bms.com> Subject: [Histonet] reduction/elimination of red cell autofluorescence on FFPE sections for IF examination To: "histonet@lists.utsouthwestern.edu" <histonet@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.thu...@bms.com 812-429-8097 ********************************************************************************* 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.vanderl...@amc.uva.nl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
