Explores the methods and applications in the expanding fluorescence field

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts

Part I: Steady-State Fluorescence Spectroscopy (SSFS). How to Collect National Institute of Standards and Technology (NIST) Traceable Fluorescence Excitation and Emission Spectra.- Steady State Fluorescence Polarization/Anisotropy for the Study of Protein Interactions.- Quantitative Fluorescence Spectral Analysis of Protein Denaturation.- High-Pressure Fluorescence Applications.- Part II: Time-Resolved Fluorescence Spectroscopy (TRFS). Frequency Domain Fluorometry: Theory and Application.- Polar Plot Representation of Time-Resolved Fluorescence.- Ensemble and Single Molecule Detected Time Resolved FRET Methods in Studies of Protein Conformations and Dynamics.- MD + QM Correlations with Tryptophan Fluorescence Spectral Shifts and Lifetimes.- Analysis of Time-Dependent Red Shifts in Fluorescence Emission from Tryptophan Residues in Proteins.- Global Analysis of Time-Resolved Fluorescence Data.- Nanometrology.- Upconversion Spectrophotofluorometry.- Subpicosecond Kerr-Gate Spectrofluorometry.- Photo-Induced Electron Transfer Modeling to Simulate Flavoprotein Fluorescence Decay.- Part III: Fluorescent Probe Development (FPD). Biosynthetic Incorporation of Trp Analogs in Proteins.- Optimization of Fluorescent Proteins.- Monitoring Membrane Properties and Apoptosis Using Membrane Probes of the 3-Hydroxyflavone Family.- Part IV: Fluorescence Microscopy: Fluorescence Recovery After Photobleaching (FRAP). Rectangle FRAP for Measuring Diffusion with a Laser Scanning Microscope.- Part V: Fluorescence Microscopy: Förster Resonance Energy Transfer Imaging (FRETim). A Quantitative Protocol for Intensity-Based Live Cell FRET Imaging.- Part VI: Fluorescence Microscopy: Fluorescence Lifetime Imaging(FLIM). Widefield Fluorescence Lifetime Imaging with Multi-Anode Detectors.- Global Analysis of FRET- FLIM Data in Live Plant Cells.- Time-Resolved Fluorescence Anisotropy Imaging.- Multi-Modal Fluorescence Imaging Spectroscopy.- Part VII: Fluorescence Microscopy: Fluorescence Fluctuation Spectroscopy (FFS). Application of Fluorescence Correlation Spectroscopy (FCS) to Measure the Dynamics of Fluorescent Proteins in Living Cells.- Fluorescence Cross-Correlation Spectroscopy (FCCS) in Living Cells.- Quantifying Lipid-Protein Interaction by Fluorescence Correlation Spectroscopy (FCS).- PET-FCS: Probing Rapid Structural Fluctuations of Proteins and Nucleic Acids by Single-Molecule Fluorescence Quenching.- Z-Scan Fluorescence Correlation Spectroscopy as a Tool for Diffusion Measurements in Planar Lipid Membranes.- Scanning Fluorescence Correlation Spectroscopy (SFCS) with a Scan Path Perpendicular to the Membrane Plane.- Implementation and Application of Pulsed Interleaved Excitation for Dual-Color FCS and RICS.- Quantitative Study of Protein-Protein Interactions in Live Cell by Dual Color Fluorescence Lifetime Cross-Correlation Spectroscopy.- Brightness Experiments.- Global Analysis of Autocorrelation Functions and Photon Counting Distributions in Fluorescence Fluctuation Spectroscopy.- Simulation of Autocorrelation Function and Photon Counting Distribution in Fluorescence Fluctuation Spectroscopy.- Part VIII: Fluorescence Microscopy: Single-Molecule Fluorescence Spectroscopy (smFS). Single-Molecule Fluorescence of Nucleic Acids.- Photoswitchable Fluorescent Proteins for Superresolution Fluorescence Microscopy Above the Diffraction Limit of Light.