Intracellular pH and BCECF
colette charland (ccharlan@zoo.uvm.edu)
Fri, 11 Oct 1996 14:14:16 -0500
I've read Dr. Hedley recent publication in Cytometry regarding
measurements of intracellular pH, which was quite helpful to a "new
investigator" but additionally pointed me in the right direction in terms of
setting up the flow. However I am still unsure about whether I am looking
at the ratio correctly. It is the first time that I am involved in this
type of analysis so any references or guidance is greatly appreciated.
At least on this first run, it appears that only a select population
of white blood cells pick up the BCECF. Consequently what it has left me
with is a distinctly positive peak as well as a negative region at 525 and
some low level staining at 640. My question is when setting up the ratio
between the two signals, should I be looking at the entire fluorescent
signal over all channels, or just at what is an obvious positive signal?
Secondly, is there any chance of using this reagent along with a cell
surface marker tagged with PE?
If anyone knows of a few good references to get me going, I'm glad
to do my homework. Thanks in advance for any replies.
Colette Charland
University of Vermont
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