On Mon, 28 Aug 1995 20:19:29 EDT, Judy Drazba wrote:
 
>Can someone out there can help me get rid of, or at least reduce the
>autofluorescence from blood vessels in our sections.  We do all of our
>imaging on brain slices (10-100 microns) fixed in 4% Paraform./PBS. I've
>tried various blocking buffers and routinely include glycine to help quench
>aldehydes to no avail.  Any suggestions??
 
If the autofluoresence is coming from the elastin fibres in the blood
vessel wall, try incubating your labelling sections in 0.5% Pontamine sky
blue 5BX (C.I. 24400, BDH Laboratories, Gurr) in buffer or saline for 10
minutes followed by a 5 minute wash before mounting. It appears that this
shifts the usual green autofluorescence emission of elastin into a red
emission. This works well in lung tissue using fluorescein labelled probes
but not so well for rhodamine or Texas Red labelled preparations.
Good luck.
Paul Rigby
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Dr Paul Rigby
Biomedical Confocal Microscopy Research Centre
Department of Pharmacology                   Telephone:  61-9-3462819
University of Western Australia                    Fax:  61-9-3463469
Nedlands WA 6907
Australia                     Email: [log in to unmask]
 
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