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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