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 --------------------------------------------------------------------- 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] ---------------------------------------------------------------------