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>
> We have been running some pH studies using SNARF-1 on myocytes. The
> investigator wants to use Nigericin to calibrate the pH measurments but we have
> some concern about contaminating the sample lines and flow cell in the flow
> cytometer. A very recent paper by P.H. Richmond & R.D. Vaughan-Jones (Pflugers
> Arch - Eur J Physiol, No. 254, 1997- I have only provisional page numbers)
> indicates that residual nigericin can strongly affect subsequent nigericin-free
> samples in a superfusion flow chamber. They developed several extensive
> cleaning procedures to remove the contaminating nigericin from their apparatus.
> I have searched through the flow literature and have found references in
> which nigericin is mentioned as a means of calibrating measurements, but have
> found nothing addressing contamination of the instrument or cleaning nigericin
> from tubing/flow cells. Does anyone have experience with this compound and is
> contamination a problem in a flow cytometric application?
>
> --
>
> Wayne Green
> wgreen@genetics.utah.edu
Wayne, we have used nigericin extensively in the past to calibrate a
Coulter EPICS V flow cytometer for measuring intracellular pH with
ADB or SNARF. We have not experienced any problems with residual
nigericin.
Mike Fox
Chairman, Cell and Molecular Biology Graduate Program
Dept. of Radiological Health Sciences mfox@cvmbs.colostate.edu
Colorado State University Ph: (970)491-7618
Ft. Collins, CO 80523 USA Fax:(970)491-0623
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web