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Proceedings Paper

Advances in polarized fluorescence depletion measurement of cell membrane protein rotation
Author(s): B. George Barisas; N. A. Rahman; Thomas Londo; J. R. Herman; Deborah A. Roess
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Paper Abstract

The laser microscopic method of polarized fluorescence depletion (PFD) has permitted the rotational dynamics of functional membrane proteins to be measured on single cells under physiological conditions. This provides information comparable to that obtained by the older technique of time-resolved phosphorescence anisotropy (TPA) but with a greater than 1000- fold reduction in sample requirements. A new cuvette implementation of PFD methods, together with a data analysis model appropriate to this new experimental geometry, permits small volumes of dilute cell suspensions to be examined in their entirety. Data can thus be obtained at rates comparable to TPA. The instrument can also be used for TPA measurements and this permits direct comparison of PFD and TPA results obtained under otherwise identical conditions. In addition, a new photomultiplier gating device and system timing strategy reduce the minimum observable rotational correlation times to < 2 microsecond(s) ec, a 10-fold improvement over previous systems. These speed improvements have been examined in studies of BSA rotation in glycerol solution.

Paper Details

Date Published: 1 June 1991
PDF: 12 pages
Proc. SPIE 1432, Biomolecular Spectroscopy II, (1 June 1991); doi: 10.1117/12.44208
Show Author Affiliations
B. George Barisas, Colorado State Univ. (United States)
N. A. Rahman, Colorado State Univ. (United States)
Thomas Londo, Colorado State Univ. (United States)
J. R. Herman, Colorado State Univ. (United States)
Deborah A. Roess, Colorado State Univ. (United States)

Published in SPIE Proceedings Vol. 1432:
Biomolecular Spectroscopy II
Robert R. Birge; Laurence A. Nafie, Editor(s)

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