Simultaneous measurement of circular dichroism and fluorescence polarization anisotropy

John Clark Sutherland

doi:10.1117/12.469782

4 June 2002 Simultaneous measurement of circular dichroism and fluorescence polarization anisotropy

John Clark Sutherland

Proceedings Volume 4625, Clinical Diagnostic Systems: Technologies and Instrumentation; (2002) https://doi.org/10.1117/12.469782
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

Citation Download Citation

John Clark Sutherland "Simultaneous measurement of circular dichroism and fluorescence polarization anisotropy", Proc. SPIE 4625, Clinical Diagnostic Systems: Technologies and Instrumentation, (4 June 2002); https://doi.org/10.1117/12.469782

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