Re: endogenous staining in rats


Also -
Block with H202 prior to  the secondary rather than before the primary antibody.  Makes a big difference.

Jacqueline M. O'Connor HT(ASCP)
Abbott Laboratories
Global Pharmaceutical Research and Development
Discovery Chemotheraputics
Fax 847.938.3266

Geoff McAuliffe <>

04/22/2003 02:32 PM

        To:        "Featherstone, Annette" <AFeatherstone@KaleidaHealth.Org>
        cc:        "'Gayle Callis'" <>,
        Subject:        Re: endogenous staining in rats

Get a fresh bottle of H2O2. Hydrogen peroxide turns into H2O over time.
I get a fresh bottle of 30 H2O2 every year, no exceptions.


Featherstone, Annette wrote:

>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 []
>Sent: Tuesday, April 22, 2003 13:30
>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)

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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