Time-Resolved Fluorescence Spectroscopy in Biochemistry and Biology (Nato Science Series A:, 69)

At the time that the editors conceived the idea of trying to organize the meeting on which the contents of this volume are based and which became, in March 1980, a NATO Advanced Study Institute, the techniques of time-resolved fluorescence spectroscopy, in both the nanosecond and sub-nanosecond time-domains, might reasonably have been said to be coming of age, both in their execution and in the analysis and interpretation of the results obtained. These techniques, then as now, comprised mainly a number of pulse methods using laser, flash-lamp or, most recently, synchrotron radiation. In addition, significant developments in the more classical phase approach had also rendered that method popular, utilizing either modulation of an otherwise continuous source or, again recently, the ultra-rapid pulse rate attainable with a synchrotron source. In general terms, time-resolved fluorescence studies are capable, under appropriate conditions, of supplying direct kinetic information on both photophysics and various aspects of molecular, macromolecular and supramolecular structure and dynamics. The nanosecond and sub-nanosecond time-scales directly probed render these techniques particularly appropriate in studying relaxation and fluctuation processes in macromolecules, particularly biopolymers (e. g. proteins, nucleic acids), in supramolecular assemblies such as cell membranes, and in a variety of relatively simpler model systems. Front Matter....Pages i-xiv Old and New Developments in Fluorescence Spectroscopy....Pages 1-20 Front Matter....Pages 21-21 Technique of Pulse Fluorometry....Pages 23-57 Phase and Modulation Fluorometry....Pages 59-80 Synchrotron Radiation as a Source to Study Time-Dependent Phenomena....Pages 81-107 Remarks on a Novel Method for Fluorescence Lifetime Measurements....Pages 109-111 Front Matter....Pages 113-113 The Pile-Up Problem in Pulse Fluorometry....Pages 115-127 Least Squares Methods of Analysis....Pages 129-141 Least Squares Methods of Analysis....Pages 143-153 A Critical Appraisal of Analytical Methods....Pages 155-168 Estimators of Exponential Decay — A Postscript....Pages 169-172 Beyond Chi-Square: Evaluation of Parametric Models Used in the Analysis of Data from Fluorescence Decay Experiments....Pages 173-195 Fluorescence Decay Analysis by the Method of Moments....Pages 199-222 Remarks on the Method of Moments for Fluorescence Decay Analysis....Pages 223-231 Systems Analysis Approaches....Pages 233-238 Fourier Transform Analysis....Pages 239-257 Analysis of Pulse Fluorometry Data by Laplace Transforms....Pages 259-270 Modulating Functions — A Deconvolution Method....Pages 271-283 An Analysis of Errors in the Phase-Plane Method of Deconvoluting Luminescence Lifetimes....Pages 285-296 Front Matter....Pages 297-297 Kinetics of Fluorescence Decay: An Overview....Pages 299-317 Excited-State Proton Transfer....Pages 319-340 Front Matter....Pages 297-297 Kinetic Aspects of Exciplex Photophysics....Pages 341-362 The Influence of Diffusion on Fluorescence Quenching....Pages 363-392 Solvent Relaxation....Pages 393-410 Long-Range Nonradiative Transfer of Electronic Excitation Energy....Pages 411-450 Energy Migration in Disordered Systems....Pages 451-461 Convolution Kinetics....Pages 463-480 Front Matter....Pages 481-481 Measurement of Brownian Correlation Times by Pulse Fluorometry....Pages 483-495 Fluorescence Anisotropy Decay and Brownian Rotational Motion: Theory and Application in Biological Systems....Pages 497-521 Energy Migration and Fluorescence Depolarization: Structural Studies of Ethidium Bromide-Nucleic Acid Complexes....Pages 523-539 Theory and Application of Differential Polarized Phase Fluorometry....Pages 541-554 Membrane Structure and Dynamics by Fluorescence Probe Depolarization Kinetics....Pages 555-604 Front Matter....Pages 613-613 Comments on the Tryptophan Lifetime Puzzle....Pages 615-620 The Excited-State Deprotonation of the Indole N-H of Tryptophan....Pages 621-622 A Preliminary Application of Phase/Modulation Fluorometry to the Tryptophan Problem....Pages 623-627 The Photophysics of Tryptophan....Pages 629-633 The Fluorescence Decays of Tryptophan in Solution at Neutral pH and in Horse Liver Alcohol Dehydrogenase....Pages 635-643 Time-resolved Spectroscopy of Tryptophyl Fluorescence in Proteins: Application to Yeast 3-Phosphoglycerate Kinase....Pages 645-649 Intrinsic Fluorescence of Proteins....Pages 651-725 Fluorescence in Aqueous Micellar Solutions....Pages 729-737 Energy Migration in Photosynthesis....Pages 741-754 Front Matter....Pages 613-613 Time-Resolved Fluorescence and Photosynthesis....Pages 755-765 Epilogue....Pages 767-768 Back Matter....Pages 769-785