I would like to introduce myself and CRI to this group using as an excuse
this query about overlapping dyes. If the question relates to microscopy as
opposed to flow cytometry (which I tend to doubt), then my answer will have
some relevance.
Briefly, my company makes tunable spectral filters that allow one to take a
number of images at different wavelengths (the bandpass can be as narrow as
5 nm, and the bandpass can be tuned with 1-nm precision; the spectral range
is typically from 450 or so to 720 nm for our visible-range filters). With
the spectral information, it is possible to unmix signals from dyes that
overlap substantially (peak to peak distances of 5 to 10 nm or less), with
overlap in the rest of the spectral range as well. We have used it to
separate greenish autofluorescence from Cy2 green dye emissions very
sucessfully. In short, for microscopy applications using the dyes Elena
lists, we should be able to unmix the signals readily without change in
labeling technique.
I would be happy to discuss this topic more online or off.
Thanks,
Richard
Richard Levenson, MD
Research Director, Biomedical Systems
CRI, Inc.
35-B Cabot Rd.
Woburn, MA 01801
781-935-9099 x 204 781-935-3388 fax
rlevenson@cri-inc.com www.cri-inc.com
----- Original Message -----
From: "Elena Soriano" <E.Soriano@iam.boku.ac.at>
To: cyto-inbox
Sent: Monday, May 28, 2001 7:45 AM
Subject: to stain RNA under GFP presence
I please for an advice about what fluorochromes could I use to stain RNA in
presence of GFP. For RNA I allways used pyronyne-Y, but in this case its
fluorescence is strongly overlapped by GFP. I have the same trouble with
FITC for total protein content.
Does someone knows what alternatives could I have and where could I
purchase them?
Thank you.
Elena.
-----------------------------------------
Elena Soriano
c/o Institut für Angewandte Mikrobiologie
Univ.f.Bodenkultur Wien
Tel: +43 1 36006 6241
Fax: +43 1 36 97 615
http://www.boku.ac.at/iam
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