endogenous staining in rats

From:"Featherstone, Annette"

Help. I am having trouble with endogenous peroxidase staining in my rat
tissues. Even though I am blocking with H2O2 in methanol, it still appears
as though I am getting blood cell staining. Any suggestions?

-----Original Message-----
From: Gayle Callis [mailto:gcallis@montana.edu]
Sent: Tuesday, April 22, 2003 13:30
To: histonet@pathology.swmed.edu
Subject: confocal and paraffin sections


There is a confocal listserver that is superb!
CONFOCAL@LISTSERV.BUFFALO.EDU - they can also help you with special CLSM

There is review article on autofluorescence, but have it at home for some
light reading, so will resend on title. 

Access this article, Control of Autofluorescence of Archival
Formaldehyde-fixed, Paraffin-embedded Tissue in Confocal Laser Scanning
Microscopy (CLSM) 
Werner Baschonga,b, Rosmarie Suetterlina, and R. Hubert Laengc 

Depending on Confocal laser scanning microscope, one can actually separate
GFP fluorescence from autofluorescence with the Zeiss CLSM 510 META using
the Lambda mode if you have that setup. This may also be the case with
certain other fluorochromes using this tidy little detection system. The
Lambda detector aka mode, using ROI can be used to get rid of
autofluorescence.  I saw this in use, it was wonderful. 

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis@montana.edu

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