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During the past decade, fluorescence techniques have come to occupy a position of central importance in biochemistry. Such areas as laser techniques, radiation less energy transfer, and nanosecond fluorometry have evolved from esoteric research specialties into standard procedures that are applied routinely to bio chemical problems. Indeed, discussion of the above three areas occupies the greater part of this book. Its level and approach are appropriate for the bio logical or physical scientist who is interested in applying fluorescence techniques, but is not necessarily an expert in this area. The coverage of the literature has, in general, been selective rather than exhaustive. It is likely that what is summarized here will prove resistant to the erosion of time and provide a basis for the future evolution of this rapidly developing area of science. Robert F. Steiner Catonsville, Maryland vii Contents Chapter 1 Some Principles Governing the Luminescence of Organic Molecules R. M Hochstrasser 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2. Spontaneous Emission . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2. 1. General Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2. 2. Luminescence from Nearby States . . . . . . . . . . . . . . . . . . . . . . . . . 4 2. 3. Multiple State Decay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3. Molecular Luminescence Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . 5 3. 1. The Transition Dipole Moment . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 3. 2. Determination of Transition-Moment Directions from Fluorescence . . . 8 3. 3. Polarization of Fluorescence fromCrystals. . . . . . . . . . . . . . . . . . . . 9 4. Principles of Luminescence Experiments Carried Out with Lasers. . . . . . . 12 4. 1. Nonlinear Processes and Optical Pumping . . . . . . . . . . . . . . . . . . . 12 5. Coherent Interactions of Molecules and Light . . . . . . . . . . . . . . . . . . . . 16 5. 1. The Distinctions between Fluorescence and Resonance Raman Effects 17 6. Ultrafast Fluorescence Decay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table des matières
1 Some Principles Governing the Luminescence of Organic Molecules.- 1. Introduction.- 2. Spontaneous Emission.- 3. Molecular Luminescence Characteristics.- 4. Principles of Luminescence Experiments Carried Out with Lasers.- 5. Coherent Interactions of Molecules and Light.- 6. Ultrafast Fluorescence Decay.- 7. The Effects of Inhomogeneous Distributions.- 8. Bibliography.- 2 Covalent Fluorescent Probes.- 1. Introduction.- 2. Primary Considerations in Fluorescent Labeling of Biomolecules.- 3. Covalent Labeling of Biomolecules.- 4. Selective Modification Reactions.- 5. References.- 3 Nanosecond Pulse Fluorimetry of Proteins.- 1. Introduction.- 2. Instrumentation and Data Analyses.- 3. Fluorescence Lifetime Studies.- 4. Fluorescence Quenching Studies.- 5. Fluorescence Energy Transfer for Distance Measurements in Proteins.- 5. Nanosecond Pulse Fluorimetry Studies of Muscle Contractile Proteins.- 7. References.- 4 The Use of Fluorescence Anisotropy Decay in the Study of Biological Macromolecules.- 1. Introduction.- 2. Theory.- 3. Experimental Procedures: Measurement of Anisotropy Decay.- 4. Applications of Fluorescence Anisotropy.- 5. References.- 5 Plasma Lipoproteins: Fluorescence as a Probe of Structure and Dynamics.- 1. Introduction.- 2. Structural Studies of Native Lipoproteins, Apoproteins, and Reassembled Lipoproteins, Using Intrinsic Protein Fluorescence and Covalently Attached Fluorescence Probes.- 3. Extrinsic Fluorescence Probes of Lipoprotein Structure and Function.- 4. Dynamics of Lipid Transfer.- 5. Summary and Perspectives.- 6. References.- 6 Fluorescent Dye-Nucleic Acid Complexes.- 1. Introduction.- 2. Intercalating and Nonintercalating Dyes.- 3. Nucleic Acid-Dye Binding Isotherms.- 4. Fluorescence Lifetimes and Quantum Yields.- 5. Decay of FluorescenceAnisotropy.- 6. Radiationless Energy Transfer.- 7. Cytological Applications.- 8. References.
