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.
Original language | English (US) |
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Pages (from-to) | 126-136 |
Number of pages | 11 |
Journal | Proceedings of SPIE - The International Society for Optical Engineering |
Volume | 4625 |
DOIs | |
State | Published - 2002 |
Externally published | Yes |
Event | Clinical Diagnostic Systems: Technologies and Instrumentation - San Jose, CA, United States Duration: Jan 22 2002 → Jan 24 2002 |
Keywords
- Circular dichroism
- Fluorescence polarization anisotropy
- Photoelastic modulator
- Stopped flow kinetics
- Ultraviolet absorption spectra
ASJC Scopus subject areas
- Electronic, Optical and Magnetic Materials
- Condensed Matter Physics
- Computer Science Applications
- Applied Mathematics
- Electrical and Electronic Engineering