Re: Wavelength Shift Calibration

Robert F. Murphy (MURPHY@A.CFR.CMU.EDU)
Mon, 13 Jul 1992 15:44:38 EDT

In response to Eric Martz's mail message regarding use of beads
for calibration of ion measurements by flow cytometry, the
first use of that method that I am aware of is described in
the paper "Endosome pH measured in single cells by dual fluorescence
flow cytometry: Rapid acidification of insulin to pH 6" (1984)
J. Cell Biol. 98:1757-1762 (see Figure 1).

For calibration of illuminated volume by a similar method and
a general discussion of discrimination of bound and free ligand
by flow, see Chapter 18 of Flow Cytometry and Sorting, Second Edition,
(Melamed, Lindmo, Mendelsohn, eds.) Wiley-Liss, Inc., 1990, pp. 355-366.
There is also a discussion of the possible effect of AC-coupling
circuits.

Bob Murphy
Carnegie Mellon University
murphy@a.cfr.cmu.edu

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