Re: Texas Red and RBCs

From:Gayle Callis

The March, 2002 J of Histochemistry and Cytochemistry had a wonderful
article on quenching autoflourescence.  Go to www.JHC.com and look it up.  

At 11:48 AM 2/27/02 -0500, you wrote:
>I need some help with an autofluorescence problem.  I'm using immersion
>fixed FFPE rat cardiac tissue and have been trying a Texas Red
>conjugated secondary antibody.  When I look at the tissues using either
>a Texas Red or a WG filter cube, the red blood cells light up like
>christmas.  I'm using a connexin antibody, so the RBCs should not be
>positive.  The fluorescence doesn't appear to be due to antibody
>nonspecificity as I've seen this in unstained tissues as well.  I've
>tried to quench the autofluorescence with NaBH4 (0.1% in PBS), but to no
>avail.  I'm not sure what else to try and am ready to switch to another
>fluor, but would really like to know what is going on here.  Has anyone
>else had similar problems.  Is there a way to correct this, or is Texas
>Red just not compatable with bloody tissues?
>
>
>Thanks,
>
>Brandon Schanbacher
>--
>Division of Pharmacology
>Ohio State University
>
>
>
>
>
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)





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