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Introduction to Fluorescence


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Table of Contents

What Is Fluorescence and How Is It Used?
A Nano-History of Fluorescence
General Concepts
And the Rest

Absorption of Light
Electromagnetic Radiation: Characterization
Relevant Wavelength Range
Absorption of Light by Molecules
Effect of Conjugation on Absorption
Effects of Molecular Environment on Absorption
Practical ConsiderationsInstrumentation
Light Sources
Optical Filters
Miscellaneous TopicsEmission and Excitation Spectra
Emission Spectra
Excitation Spectra
Delayed Fluorescence
Advanced Scanning MethodsPolarization and Anisotropy
Basic Principles
Some Applications of Polarization/Anisotropy
Depolarization via FRETTime-Resolved Fluorescence
Excited State Lifetimes
Instrumentation Details
Weber's Algorithm
Time Domain versus Frequency Domain
Magic Angles
Anisotropy Decay/Dynamic Polarization
Dynamic Polarization
PhasorsQuantum Yields and Quenching
Determination of QYs
Kasha-Vavilov Rule
Quenching and Membrane SystemsFoerster Resonance Energy Transfer
Basic Principles
How Do We Determine the Efficiency of Energy Transfer (E)?
The Orientation Factor
How Do We Determine 2?
Homo-Transfer of Electronic Excitation Energy
Examples of FRET Applications
Advanced TreatmentsBrief Overview of Fluorescence Microscopy
Introduction of Fluorophores into Living Cells
Fluorescence Microscopy Approaches
Specialized TechniquesFluorophores
Where Do Fluorophores Come From?
How Does One Choose a Fluorophore?
Intrinsic Probes
Extrinsic Probes
Labeling Proteins In Vitro
Membrane Probes
Nucleic Acid Probes and Nucleotide/Nucleoside
Ion Probes
pH Probes
Molecular Beacons
Voltage-Sensitive Dyes
Miscellaneous Probes
Fluorescent ProteinsIntrinsic Protein Fluorescence
Electronic Energy Transfer in Proteins
Use of Site-Directed Mutagenesis
Tryptophan Analogs
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.

About the Author

University of Hawaii, Manoa, USA


"An essential contribution to educating scientists in the principles of fluorescence. It will also be an important addition to the libraries of practitioners applying the principles of molecular fluorescence."
-Ken Jacobson, Kenan Distinguished Professor of Cell Biology and Physiology, University of North Carolina at Chapel Hill

"An exquisite compendium of fluorescence and its applications in biochemistry enriched by a very exciting historical perspective. This book will become a standard text for graduate students and other scientists."
-Drs. Zygmunt (Karol) Gryczynski and Ignacy Gryczynski, University of North Texas Health Science Center

"... truly a masterwork, combining clarity, precision, and good humor. The reader, novice or expert, will be pleased with the text and will not stop reading. It is a formidable account of the fluorescence field, which has impacted the life sciences so considerably in the last 60 years."
-Jerson L. Silva, M.D., Ph.D., Professor and Director, National Institute of Science and Technology for Structural Biology and Bioimaging, Universidade Federal do Rio de Janeiro

"Dr. Jameson's book fills a gap in the literature available for the student approaching the applications of fluorescence techniques as well as for the researcher practicing them daily in the laboratory setting. The subject is presented by the author in a clear didactic manner; the anecdotes and historical references make the book truly a delight to read."
-Dr. Beniamino Barbieri, President, ISS Inc.

"An excellent textbook! This comprehensive overview written at an introductory level covers fundamental aspects, principles of instrumentation, and practical applications of biological fluorescence, while providing many valuable tips. In essence, this book reflects the excellent didactic skills and vast experience of the author in research and teaching fluorescence. Highly recommendable for undergraduate teaching. Also excellent material for active researchers who are learning to apply fluorescence methods."
-Prof. Luis Bagatolli, Center for Biomembrane Physics, Department of Biochemistry and Molecular Biology, University of Southern Denmark

"I am very impressed by the author's lucid treatment. This book is a `must read' for students and researchers."
-Prof. Amitabha Chattopadhyay, Ph.D., FRSC, J.C. Bose Fellow, Outstanding Scientist, Centre for Cellular & Molecular Biology, Hyderabad

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