Re: [Histonet] 2) Immunofluorescence and background

In a message dated 3/31/2005 5:11:55 PM US Mountain Standard Time, writes:

> have a different but similar problem with Immunofluorescence and
> background. I'm cutting rat spinal cord sections that have been perfused and
> post fixed in 4% paraformaldehyde, cryoprotected and cut as frozen sections.

Hi Cath,

Yes, it's almost certainly autofluorescence due to the aldehyde 
fixation...perhaps you can cut down on the fluorescence by shortening the fixation time, or 
you can try using sodium borohydride treatment on the sections before 
beginning the staining protocol, or perhaps quenching the reactive groups with amines 
(like lysine.)

The subject has been discussed a few times on Histonet.  You can search the 
archives by going to:

Use the search terms "autofluorescence paraformaldehyde" (w/o quote marks) to 
see what's been said about the problem.



Robert (Bob) Chiovetti, Ph.D.
Independent Consultant for The Science, Technology and Industrial Sectors
132 North Elster Drive
Tucson, AZ 85710-3212 USA
Tel./Fax 520-546-4986
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