More: autofl.

<< Previous Message | Next Message >>
From:"Karen D. Larison" <> (by way of histonet)
To:histonet <>
Content-Type:text/plain; charset="us-ascii"

Out of curiosity, I just looked at some Bouin's-fixed lamprey muscle under my
fluorescent scope and it wasn't autofluorescent in my rhodamine,
fluorescein or
DAPI filter sets.  So I don't think the picric acid in your fix is the
In fact, one might worry that the picric acid would instead quench the
fluorescence of your signal, particularly if it imparts a yellow color to the
tissue.  In general, chromogens and metals tend to quench fluorescence.

Karen Larison

Date:          Tue, 22 Dec 1998 08:54:56 -0500
From:          "Michael J. Lyon, Ph.D." <lyonm@VAX.CS.HSCSYR.EDU>
Subject:       Autofluorescence
To:            "Histonet (E-mail)" <>


I have just started doing immunostaining of muscle dissections.  The tissue
been immersion fixed in 4% paraformaldehyde 0.2% Picric acid for about 24 hrs
at 4C.  My primary antibodies are all fa
rly dilute at 1:3000 or 1:4000, secondary is at 1:400 and I am using CY3 at
1:600.  I have tried ammonium chloride and sodium borohydride but still have
considerable background from the muscle fiber
.  I was thinking of switching to a different fixation.  Any suggestions would
be appreciated.  Hope I have given sufficient info.

Thanks in advance


<< Previous Message | Next Message >>