Introduction to Fluorescence

✍ Scribed by Jameson, David M

• Publisher: CRC Press
• Year: 2014
• Tongue: English
• Leaves: 308
• Category: Library

✦ Synopsis

"A concise introduction to fundamental principles, this book presents the most important aspects of fluorescence spectroscopy, particularly as they are applied to biology, biochemistry, and biophysics. The author focuses on the essential aspects of fluorescence to encourage readers to design experiments, begin their own fluorescence measurements, and access the literature with greater comprehension. After a general definition of the field and an overview of the nature of light and absorption by biomolecules, the text covers instrumentation, emission spectra, excitation spectra, polarization, anisotropy, quenching, time-resolved methods, and FRET"-- Read more...

Abstract: "A concise introduction to fundamental principles, this book presents the most important aspects of fluorescence spectroscopy, particularly as they are applied to biology, biochemistry, and biophysics. The author focuses on the essential aspects of fluorescence to encourage readers to design experiments, begin their own fluorescence measurements, and access the literature with greater comprehension. After a general definition of the field and an overview of the nature of light and absorption by biomolecules, the text covers instrumentation, emission spectra, excitation spectra, polarization, anisotropy, quenching, time-resolved methods, and FRET"

✦ Table of Contents

Content: Introduction What Is Fluorescence and How Is It Used? A Nano-History of Fluorescence General Concepts And the Rest Absorption of Light Electromagnetic Radiation: Characterization Photons Relevant Wavelength Range Absorption of Light by Molecules Effect of Conjugation on Absorption Effects of Molecular Environment on Absorption Practical Considerations Instrumentation Light Sources Optical Filters Monochromators Polarizers Detectors Miscellaneous Topics Emission and Excitation Spectra Emission Spectra Excitation Spectra Delayed Fluorescence Advanced Scanning Methods Polarization and Anisotropy Basic Principles Some Applications of Polarization/Anisotropy Depolarization via FRET Time-Resolved Fluorescence Excited State Lifetimes Instrumentation Details Weber's Algorithm Time Domain versus Frequency Domain Magic Angles Anisotropy Decay/Dynamic Polarization Dynamic Polarization Phasors Quantum Yields and Quenching Determination of QYs Kasha-Vavilov Rule Quenching Quenching and Membrane Systems Forster Resonance Energy Transfer Background Basic Principles How Do We Determine the Efficiency of Energy Transfer (E)? The Orientation Factor How Do We Determine kappa2? Homo-Transfer of Electronic Excitation Energy Examples of FRET Applications Advanced Treatments Brief Overview of Fluorescence Microscopy Introduction of Fluorophores into Living Cells Fluorescence Microscopy Approaches Specialized Techniques Fluorophores Where Do Fluorophores Come From? How Does One Choose a Fluorophore? Intrinsic Probes Extrinsic Probes Photostability Labeling Proteins In Vitro Membrane Probes Nucleic Acid Probes and Nucleotide/Nucleoside Analogs Biosensors Ion Probes pH Probes Molecular Beacons Voltage-Sensitive Dyes Miscellaneous Probes Fluorescent Proteins Intrinsic Protein Fluorescence Spectra Polarization Lifetime Electronic Energy Transfer in Proteins Use of Site-Directed Mutagenesis Tryptophan Analogs Applications Protein Unfolding/Refolding Protein-Protein and Protein-DNA Interactions Tyrosine Studies Phasors and Protein Fluorescence Appendix: Rogue's Gallery of Fluorescence Artifacts and Errors Index Additional Reading appears at the end of each chapter