[Histonet] Re: DIF tissue in GLUT

WILLIAM DESALVO wdesalvo.cac <@t> hotmail.com
Thu Aug 27 12:51:21 CDT 2009

I suggest you use a counterstain for your IF to reduce the autofluoresence. Evans Blue - the product can be used as a counterstain in immunohistochemistry when using FITC. After staining for immunofluorescence, dip sections in a 0.1% (w/v) in water solution of Evans Blue for 5-10 minutes. Rinse well in fresh PBS or water before coverslipping. Reference: Immunocytochemistry, Theory and Practice, p. 82 (1988). Purchase from Sigma-Aldrich.

William DeSalvo, B.S., HTL(ASCP)

> From: TJJ <@t> stowers.org
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Thu, 27 Aug 2009 12:18:47 -0500
> Subject: [Histonet] Re: DIF tissue in GLUT
> Anne,
> Tissue that has been fixed in glutaraldehyde has very, VERY bright autofluorescence. Unless there is some way to minimize this (none that I'm aware of), your immunofluorescence will be impossible to read over the background signal.
> Teri Johnson, HT(ASCP)QIHC
> Managing Director Histology Facility
> Stowers Institute for Medical Research
> 1000 E. 50th St.
> Kansas City, MO 64110
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