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Proceedings Paper

Simultaneous measurement of circular dichroism and fluorescence polarization anisotropy
Author(s): John Clark Sutherland
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Paper Abstract

Circular dichroism and fluorescence polarization anisotropy are important tools for characterizing biomolecular systems. Both are used extensively in kinetic experiments involving stopped - or continuous flow systems as well as titrations and steady-state spectroscopy. This paper presents the theory for determining circular dichroism and fluorescence polarization anisotropy simultaneously, thus insuring the two parameters are recorded under exactly the same conditions and at exactly the same time in kinetic experiments. The approach to measuring circular dichroism is that used in almost all conventional dichrographs. Two arrangements for measuring fluorescence polarization anisotropy are described. One uses a single fluorescence detector and signal processing with a lock-in amplifier that is similar to the measurement of circular dichroism. The second approach uses classic T format detection optics, and thus can be used with conventional photon-counting detection electronics. Simple extensions permit the simultaneous measurement of the absorption and excitation intensity corrected fluorescence intensity.

Paper Details

Date Published: 4 June 2002
PDF: 11 pages
Proc. SPIE 4625, Clinical Diagnostic Systems: Technologies and Instrumentation, (4 June 2002); doi: 10.1117/12.469782
Show Author Affiliations
John Clark Sutherland, Brookhaven National Lab. and East Carolina Univ. (United States)

Published in SPIE Proceedings Vol. 4625:
Clinical Diagnostic Systems: Technologies and Instrumentation
Gerald E. Cohn, Editor(s)

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