fluorescence intensity from large cells/estimating surface area
James Leary (JLEARY@BEACH.UTMB.EDU)
Thu, 06 Apr 1995 11:07:50 -0500 (CDT)
I've spent a fair bit of time worrying about accurately estimating cell
size, surface area, shape, etc. by flow cytometry. Forward scatter is a
dangerous (and crude), nonlinear measurement for measuring cell size. It
works reasonably well for spherical billiard balls like CHO cells but very
badly for most cell types. If you have the measurement capabiity on your
instrument use pulse width time-of-flight, with the ability to subtract
out the laser beam width. For an example you might want to read the
article we published in Cyometry 6: 151-158, 1985. We made measurements of
cell diameter, surface area and volume, using it to measure transferrin
receptor density per unit surface area. The measurement does assume that
the cells are spherical and without a highly convoluted surface structure
(which may or may not be true for your cells of interest). But in general
it is a very good measurement.
-------------------------------------------------------------
James F. Leary Professor of Internal Medicine
Director, Molecular Cytometry Unit-Route 0841
University of Texas Medical Branch
Galveston, Texas 77555-0841
Tel: 409-747-1930 Fax: 409-772-6527
email: jleary@beach.utmb.edu
-------------------------------------------------------------
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
