Re: Intracellular pH

From: Elizabeth R. Simons (bu) (esimons@bu.edu)
Date: Mon Sep 30 2002 - 11:18:40 EST


BCECF works very well on a flow cytometer with 488 nm excitation; maximal
fluorescence is at 530 but it has a very broad emission and still can be
read with a 575 or a 600nm long pass filter. We have a number of papers
using it in a flow cytometer - I don't have the more recent references in
front of me but you can look them up in medline. A few of the older ones
are below (there's also one in press currently in J. Leukoc. Biol.).
Elizabeth R. Simons

A.T. Gewirtz, K.F. Seetoo, E.R. Simons. Neutrophil degranulation and
phospholipase D activation are  enhanced if the Na+/H+ antiport is blocked,
J.Leuk.Biol. 64:   98-103, 1998
E.R. Simons,  Early  Events in Receptor-mediated Neutrophil Signal
Transduction, in "Phagocyte Function: A  Guide for Research and Clinical
Evaluation; Volume I: Cytometric Cellular Analysis" (J.P. Robinson  and G.
Babcock, eds) Wiley, NY, 77-96, 1998
E.R.Simons, Flow Cytometry: Use of multiparameter kinetics to evaluate
several activation parameters  simultaneously in individual living cells.
in "Fluorescent and Luminescent Probes", 2nd Ed.Mason,  Ed.) Academic
Press, 527-539, 1999


Howard Shapiro wrote:

> Rob McCord wrote-
>
> >I'm looking for a marker to measure intracellular pH.   Practical Flow
> >Cytometry (3rd)
> >mentions both an unnamed UV excited dye used, and BCECF-AM for 488 nm
> >excitation.
> >Are these still the most widely used markers?
>
> The UV-excited dye was never that widely used; BCECF-AM is very widely
> used, and comes from Molecular Probes, as do a variety of other
> pH-sensitive dyes in the SNARF and SNAFL families. Look at the Molecular
> Probes website for references.
>
> -Howard

--
Elizabeth R. Simons, Ph.D.
Professor of Biochemistry
Boston University School of Medicine
80 E. Concord Street
Boston, MA 02118
(617) 638-4332
(617) 638-5339 FAX
esimons@bu.edu
ersimons@earthlink.net



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