An Introduction to Fluorescence Correlation Spectroscopy represents a comprehensive introduction to fluorescence correlation spectroscopy (FCS), a biophysical experimental technique increasingly used to study and quantify molecular mobility, concentrations and interactions in vitro, as well as in living cells and multicellular organisms. Students and researchers who are new to FCS can use the book as the first introduction to the technique, while those who are already using FCS regularly in their research may find it useful to deepen their understanding of the technique, its possibilities, limitations, and potential pitfalls as well as ways to avoid them. This book introduces the reader to all aspects of FCS needed for practical usage of the technique in their research. In the beginning the concept of fluorescence intensity fluctuations and their auto- and cross-correlation functions are explained to give readers an understanding of the underlying principles. This is followed by an overview of instrumental FCS setups and various ways of data collection and processing, the derivations of theoretical models relating the experimentally obtained correlation functions to the underlying molecular processes, and the description of the fitting of experimental data with those models. Mathematically more involved portions are separated from the rest of the text and can be easily skipped by readers more interested in the conceptual and practical aspects of FCS. The book contains interactive graphics and is accompanied by an interactive computable document file allowing the reader to test the dependence of FCS results on a variety of experimental parameters, and to gain practical insights into FCS data fitting. Key Features Introduces the concepts of FCS in an accessible way, supported by animations and graphics in the ebook. Includes a supplementary interactive computable document file that allows the reader to experiment with various FCS setup and fit parameters, allowing readers to test their understanding and simulate experimental outcomes. Provides rigorous mathematical derivations of fundamental FCS equations and models. Pedagogical features include questions, short reviews and critical discussions of literature relevant to the particular chapter that include applications and fundamental developments in the field of FCS.

This is the first book-length treatment of both the theoretical background to fluorescence correlation spectroscopy (FCS) and a variety of applications in various fields of science. The high spatial and temporal resolution of FCS has made it a powerful tool for the analysis of molecular interactions and kinetics, transport properties due to thermal motion, and flow. It contains an essential contribution from Nobel Prize winner M. Eigen, who is credited with inventing FCS.

An Introduction to Fluorescence Correlation Spectroscopy represents a comprehensive introduction to fluorescence correlation spectroscopy (FCS), a biophysical experimental technique increasingly used to study and quantify molecular mobility, concentrations and interactions in vitro, as well as in living cells and multicellular organisms. Students and researchers who are new to FCS can use the book as the first introduction to the technique, while those who are already using FCS regularly in their research may find it useful to deepen their understanding of the technique, its possibilities, limitations, and potential pitfalls as well as ways to avoid them. This book introduces the reader to all aspects of FCS needed for practical usage of the technique in their research. In the beginning the concept of fluorescence intensity fluctuations and their auto- and cross-correlation functions are explained to give readers an understanding of the underlying principles. This is followed by an overview of instrumental FCS setups and various ways of data collection and processing, the derivations of theoretical models relating the experimentally obtained correlation functions to the underlying molecular processes, and the description of the fitting of experimental data with those models. Mathematically more involved portions are separated from the rest of the text and can be easily skipped by readers more interested in the conceptual and practical aspects of FCS. The book contains interactive graphics and is accompanied by an interactive computable document file allowing the reader to test the dependence of FCS results on a variety of experimental parameters, and to gain practical insights into FCS data fitting. Key Features Introduces the concepts of FCS in an accessible way, supported by animations and graphics in the ebook. Includes a supplementary interactive computable document file that allows the reader to experiment with various FCS setup and fit parameters, allowing readers to test their understanding and simulate experimental outcomes. Provides rigorous mathematical derivations of fundamental FCS equations and models. Pedagogical features include questions, short reviews and critical discussions of literature relevant to the particular chapter that include applications and fundamental developments in the field of FCS.

Fluorescence methods are being used increasingly in biochemical, medical, and chemical research. This is because of the inherent sensitivity of this technique. and the favorable time scale of the phenomenon of fluorescence. 8 Fluorescence emission occurs about 10- sec (10 nsec) after light absorp tion. During this period of time a wide range of molecular processes can occur, and these can effect the spectral characteristics of the fluorescent compound. This combination of sensitivity and a favorable time scale allows fluorescence methods to be generally useful for studies of proteins and membranes and their interactions with other macromolecules. This book describes the fundamental aspects of fluorescence. and the biochemical applications of this methodology. Each chapter starts with the -theoreticalbasis of each phenomenon of fluorescence, followed by examples which illustrate the use of the phenomenon in the study of biochemical problems. The book contains numerous figures. It is felt that such graphical presentations contribute to pleasurable reading and increased understand ing. Separate chapters are devoted to fluorescence polarization, lifetimes, quenching, energy transfer, solvent effects, and excited state reactions. To enhance the usefulness of this work as a textbook, problems are included which illustrate the concepts described in each chapter. Furthermore, a separate chapter is devoted to the instrumentation used in fluorescence spectroscopy. This chapter will be especially valuable for those perform ing or contemplating fluorescence measurements. Such measurements are easily compromised by failure to consider a number of simple principles.

In color throughout, this text helps readers acquire a sound understanding of basic fluorescence theory and practice. It takes them through the history of important discoveries to the most current advances. The author introduces the fundamentals of the fluorescence phenomenon and gives detailed examples of fluorescence applications in the molecular life sciences, including biochemistry, biophysics, clinical chemistry and diagnostics, pharmaceutical science, and cell and molecular biology. The text includes references in each chapter, more than 250 figures, and the chemical structures of the most widely used fluorescent molecules.

Providing much-needed information on fluorescence spectroscopy and microscopy, this ready reference covers detection techniques, data registration, and the use of spectroscopic tools, as well as new techniques for improving the resolution of optical microscopy below the resolution gap. Starting with the basic principles, the book goes on to treat fluorophores and labeling, single-molecule fluorescence spectroscopy and enzymatics, as well as excited state energy transfer, and super-resolution fluorescence imaging. Examples show how each technique can help in obtaining detailed and refined information from individual molecular systems.