RE: Cell Volume Revisited

From: Robert C. Leif, Ph.D. (rleif@rleif.com)
Date: Sat Oct 14 2000 - 00:01:40 EST


From: Bob Leif
To: cyto-inbox

You are absolutely correct! The technology to perform electro-optical
analysis of cells has been developed. In fact, it is in the high-end Coulter
hematology analyzers. The two Coulter impedance parameters, AC (opacity) and
DC (volume)are extremely useful parameters. I might add that even in the
case of light-scatter, the commercial instruments should be set to 45 rather
than 90 degrees.

Unfortunately, I have learned that solving technical problems is certainly
more fun and usually much easier than solving people problems. In any case,
my monolithic spherical transducers and other innovations are now
off-patent. If anyone is interested in improving the bread of flow
cytometers and digital microscopes, I suggest attending the SPIE BiOS
meeting (www.spie.org, 20-26 January, 2001) including the session on
Advanced Techniques in Analytical Cytology (Thursday 25 January, 2001),
which I have the privilege to chair. If there is any interest in a means to
implement or encourage the development of modern analytical cytology
instrumentation, I will gladly organize a bull-session at the end of the
meeting. I might note that I have been honored to have as CoChairs
individuals, who have made major contributions to this field.

1. Orifice Inside Optical Elements, Coulter Electronics, R. C. Leif,
4,348,107 (1982).

2. Monitoring of a Detection Zone Utilizing Zero Order Radiation From a
Concave Reflecting Grating, Coulter Electronics, R. C. Leif, 4,351.611
(1982).

3. R. C. Leif, R. A. Thomas, T. A. Yopp, B. D. Watson, V. R. Guarino D. H.
K. Hindman, N. Lefkove and L. M. Vallarino; “Development of Instrumentation
and Fluorochromes for Automated Multiparameter Analysis of Cells”. Clin.
Chem. 23, 1492-1498 (1977).

4. R. C. Leif, S. Schwartz, C. M. Rodriguez, L. Pell-Fernandez, M. Groves,
S. B. Leif, M. Cayer, H. Crews; “Two Dimensional Impedance Studies of BSA
Buoyant Density Separated Human Erythrocytes”. Cytometry 6 pp. 13-21 (1985).

5. R. C. Leif, M. L. Cayer, W. Dailey, T. Stribling, and K. Gordon, “The use
of a Spherical Multiparameter Transducer for Flow Cytometry”. Cytometry 20,
pp 185-190 (1995).

-----Original Message-----
From: James Weaver 301-594-5879 FAX 301-594-3037
[mailto:WEAVER@CDER.FDA.GOV]
Sent: Thursday, October 12, 2000 10:55 AM
To: cyto-inbox
Subject: Re: Cell Volume Revisited
Sensitivity: Confidential



Despite what is written in various places, flow cytometry can only be
used for size measurement on homogenous particles such as plastic beads.
It does not work for particles with complex optics such as cells. With
beads, increasing size leads to increased light scatter. With cells the
situation is quite different, a treatment that resulted in a 30% change
in cell volume caused a light scatter decrease in a lymphoid cell line
and no change in light scatter from NIH3T3 cells.

	Coulter volume measurements are regarded as the gold standard of
cell volume measurements. I have done cell volume measurements on
nucleated cells in the past with no problems. If you are not seeing
changes in cell volume, than either the treatment is not causing a
change or the instrument needs repair.

-Jim Weaver


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