Presenting a detailed, hands-on approach to fluorescence spectroscopy, this book describes experiments that cover basic spectroscopy and advanced aspects of fluorescence spectroscopy. It emphasizes practical guidance, providing background on fundamental concepts as well as guidance on how to handle artifacts, avoid common errors, and interpret data. Nearly 150 experiments from biophysics, biochemistry, and the biomedical sciences demonstrate how methods are applied in practical applications. The result is a hands-on guide to the most important aspects of fluorescence spectroscopy, from steady-state fluorescence to advanced time-resolved fluorescence.
Provides a complete overview of nearly 150 experiments using fluorescence spectroscopy, from basic to advanced applications Presents laboratory methods using a variety of instrumental setups with detailed discussion of data analysis and interpretations Covers steady-state phenomena, time-resolved phenomena, and advanced methods Spans biophysical, biochemical, and biomedical applications Describes related concepts, theory, and mathematical background as well as commercially available instruments used for measurements
Chapter 1. Theory of Light and Light Interaction with Matter Basics of Light Light Polarization Interaction of Light and Chromophores Interactions Among Chromphores Scattering Summary Chapter 2. Experimental Basics Types of Cuvettes Types of Filters Measuring Transmittance/Absorbance Factors Affecting the Precision of Absorption Measurement Effect of Sample Scattering on Measured Transmission and Abosrbance Measuring Emission Efficiency of Collecting Emission: Measuring Quantum Yield Wavelength Calibration Instrucment Response Lamp Profile Peaks and Spectral Shapes Standard Deviation and Height Sums of Peaks Noise-Signal-to-Noise Ratio Sampling Rate Bandwidth Artifacts and Errors Spectral Overlap Peak Normalization Residuals Interpolation Chapter 3. Experiments Introduction Experiments Chapter 4. Fluorescence-Steady-State Phenomena Introduction Experiments Chapter 5. Steady-State Fluorescence: Applications Introduction Experiments Chapter 6. Steady-State Fluorescence Polarization: Anisotropy Introduction How to Select and Check Polarizers Transition Moments Polarized Absorption Emission Polarization (Anisotropy) Fundamental and Limiting Anistropies Consequences of Linear Transition Moments Determining the G-Factor for Spectrofluorometer Experiments Chapter 7. Fluorescence: Time-Resolved Phenomena Introduction Experiments Chapter 8. Advanced Experiments Introduction Experiments
Zygmunt (Karol) Gryczynski is a professor of molecular biology and immunology and the director of the Center for Commercialization of Fluorescence Technologies (CCFT) the University of North Texas Health Center. Ignacy Gryczynski is a professor of cell biology and genetics and the director of the Microscopy Core Facility at the University of North Texas Health Center.