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Is the autofluorescence distributed across the retina, or is it limited
to specific cell types?  Also, which wavelengths?  Retina is full of
carotenoids --especially in the photoreceptors, and they are quite
fluorescent.  We were able to overcome it by using Rhodamine
based fluorophores, since most of the background was in the green.
We also see much less autofluorescence if we use paraffin
embedments.  I assume that all of the solvents extract most of the
carotenoids.


Date sent:              Sat, 22 Nov 2003 07:03:34 -0800
Send reply to:          [log in to unmask]
From:                   Terry Wu <[log in to unmask]>
Subject:                Reduce Auto-Fluorescence
To:                     [log in to unmask]

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello,
>
> We have found that the auto-fluorescence level in rabbit retina tissue
> is very high.  The preparations are fronzen sections of 15 um.  Fixed
> with acetone.  We are wondering if anyone has any suggestions as to
> how to reduce the auto-fluorescence level before confocal imaging.
>
> Thanks in advance for your help!
>
> Terry
>
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Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail: [log in to unmask]