Cell Structure and Function by Microspectrofluorometry

ContributorsPrefaceTomas Hirschfeld-In MemoriamPart I History 1. The Origins of Modern Fluorescence Microscopy and Fluorescent Probes I. Introduction II. The First Fluorescence Microscopes III. Technical Progress IV. Advances in Biomedical Applications V. Modern Fluorescence Microscopy in Cell and Molecular Biology VI. Development of Immunofluorescence ReferencesPart II Methods 2. Microspectroscopy and Flow Cytometry 3. From Solution Spectroscopy to Image Spectroscopy I. Fluorescence Spectra II. Fluorescence Excitation Spectrum III. Fluorescence Lifetime and Yield IV. Fluorescence Polarization References 4. High-Resolution Fluorescence and Phase Microscopy in Conjunction with Micromanipulation for In Situ Study of Metabolism in Living Cells I. Introduction II. Resolution of Transmission and Fluorescence Microscopes III. Microscope Methods IV. Long-Working-Distance Condenser for Micromanipulation V. Applications of Spectroscopy to Fluorescence Microscopy VI. Instrument Design VII. Application of Photography VIII. Future Developments References 5. FRET Microscopy: Digital Imaging of Fluorescence Resonance Energy Transfer. Application in Cell Biology I. Introduction II. Theory of Fluorescence Resonance Energy Transfer III. Measurement of FRET: Data Acquisition and Analysis IV. Experimental Methods and Results V. Discussion and Future Prospects References 6. Fluorescence Scanning Instrumentation I. Introduction II. Stage Scanning Microfluorometers III. Laser Scanning Microfluorometers IV. Composition of a Laser Scanning Microscope V. Confocal Laser Scanning VI. Characteristics of Laser Scanning VII. Applications in Laser Scanning References 7. Fluorescence Microscopy in Three Dimensions: Microtomoscopy I. Introduction II. Confocal Microscopy III. Applications IV. Discussion References 8. Fluorescence Photochemical Techniques for the Study of Transport in Cytoplasm and Cytoplasmic Models I. Introduction II. Apparatus and Methodology III. Applications to Cytoplasmic Transport IV. Applications to Cytoplasmic Models In Vitro V. Fluorescence Photoactivation VI. Conclusions References 9. Principles of Frequency-Domain Fluorescence Spectroscopy and Applications to Protein Fluorescence I. Introduction II. Comparison of Time and Frequency-Domain Measurements III. Theory of Frequency-Domain Fluorometry IV. Tryptophan Fluorescence from Proteins V. 2-GHz Frequency-Domain Fluorometry VI. Additional Applications of Frequency-Domain Fluorometry VII. Future Developments References 10. The First Picosecond in Vision I. Introduction II. Picosecond Time-Resolved Fluorescence Techniques III. Picosecond Fluorescence Spectroscopy Results IV. Discussion ReferencesPart III Metabolism 11. Microspectrofluorometry of Single Living Cells: Quo Vadis I. Introduction II. Instrumentation and Methods in Microspectrofluorometry III. Biological Material IV. Spatiotemporal Organization of Cell Metabolism V. Spatiotemporal Mapping of Other Organelles: Lysosomes VI. Fluorescence Detection of Multiorganelle Complexes Associated with the Cell's Detoxification Function VII. Other Applications VIII. Conclusions References 12. Mechanism of Action of Xenobiotics: from Molecular Spectral Studies to Microspectrofluorometry of Living Cells I. Introduction II.