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Original Articles Polarization of Scatter and Fluorescence Signals in

Summary: Original Articles
Polarization of Scatter and Fluorescence Signals in
Flow Cytometry
Charles L. Asbury, Jeanne L. Uy, and Ger van den Engh*
Department of Molecular Biotechnology, University of Washington, Seattle, Washington
Received 1 November 1999; Revision Received 31 January 2000; Accepted 16 February 2000
Background: The pulses of light scatter and fluorescence
measured in flow cytometers exhibit varying degrees of
polarization. Flow cytometers are heterogeneously sensi-
tive to this polarization, depending on the light source(s),
the optical layout, and the types of mirrors and filters
used. Therefore, fluorescence polarization can affect ap-
parent intensity ratios between particles and interfere
with schemes for interlaboratory standardization.
Methods: We investigate the degree to which polariza-
tion affects common flow cytometry measurements. Our
technique for determining polarization differs from previ-
ous methods because complete distributions of intensity
versus polarization angle are measured, rather than inten-
sities at just two orthogonal polarization angles. Theoret-


Source: Asbury, Chip - Department of Physiology and Biophysics, University of Washington at Seattle


Collections: Biology and Medicine