Re: Cover flourescence ?
I'll bet the rhodamine spheres were made of polystyrene, which are
VERY soluble in xylene. Been there, done that, bought the shirt!
Date sent: Fri, 05 Apr 2002 01:03:59 -0500
From: "J. A. Kiernan"
Subject: Re: Cover flourescence ?
To: "Dianne Holmes (by way of Histonet)"
Copies to: HistoNet@pathology.swmed.edu
> "Dianne Holmes (by way of Histonet)" wrote:
> > We are using Rhodamine microspheres to label rat brains. Got great
> > labelling - - coverslipped with DPX and lost half of the signals!!!
> > HELP! Is it the xylene? Any suggestions ...
> Xylene and DPX do not abolish fluorescence of rhodamine-labelled
> proteins, which are denatured (made permanently insoluble) when a
> apecimen passes through a chemically unreactive fixative (alcohol
> etc). You are probably correct in your assumption that a solvent
> dissolved the stuff the microspheres were made of. Do you know
> what it was? If you don't, why did you use it? Even if there is
> trade secrecy about the microspheres, their maker must surely
> issue some information about solvents to avoid.
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London, Canada N6A 5C1
Atlantic Veterinary College, U.P.E.I.
550 University Ave.
Canada, C1A 4P3
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