William, thanks for the reminder, I had totally forgotten about using a counterstain to counteract AF in aldehyde fixed samples. We have some glutaraldehyde fixed cryosections of kidney I think I'll try this with. I can assure you, the autofluorescence in these samples is magnificent! I worry about using sodium borohydride because they are cryosections and I've heard the treatment can be a bit harsh on tissue.
Here's some additional information since we're on the subject: http://www.uhnresearch.ca/facilities/wcif/PDF/Autofluorescence.pdf Kind regards, Teri -----Original Message----- From: WILLIAM DESALVO [mailto:wdesalvo....@hotmail.com] Sent: Thursday, August 27, 2009 12:51 PM To: Johnson, Teri; histonet Subject: RE: [Histonet] Re: DIF tissue in GLUT 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: t...@stowers.org > To: histonet@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 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ Windows Live: Keep your friends up to date with what you do online. Find out more.<http://windowslive.com/Campaign/SocialNetworking?ocid=PID23285::T:WLMTAGL:ON:WL:en-US:SI_SB_online:082009> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
