Handbook Of Fluorescence Spectroscopy And Imaging

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.

Melding basic and clinical science, this reference provides a comprehensive overview of the roles that biophysics, photochemistry, and computational modeling play in the biomedical applications of fluorescence spectroscopy and imaging. Penned by pioneering researchers, the Handbook of Biomedical Fluorescence discusses fundamental aspects of fluorescence generation in organic molecules within tissue, theoretical and experimental views of how light propagation in tissue can be used to interpret fluorescence signals, endogenous and exogenous fluorescence agents in medical or basic research studies, and radiation transport, diffusion theory, and the Monte Carlo method.

This is a practical introduction to single molecule fluorescence experiments, the analysis of the data, and applications of the techniques to the study of biological structure and function.

During the past two decades, there has been an increasing appreciation of the significant value that lifetime-based techniques can add to biomedical studies and applications of fluorescence. Bringing together perspectives of different research communities, Fluorescence Lifetime Spectroscopy and Imaging: Principles and Applications in Biomedical Diagnostics explores the remarkable advances in time-resolved fluorescence techniques and their role in a wide range of biological and clinical applications. Broadly accessible, the book captures the state-of-the-art of fluorescence lifetime metrology and imaging and provides current perspectives on their applications to biomedical studies of intact tissues and medical diagnosis. The text introduces these techniques within the wider context of fluorescence spectroscopy and describes basic principles underlying current instrumentation for fluorescence lifetime imaging and metrology (FLIM). It also covers the wide range of methods, including single channel (point) spectroscopy, fluorescence lifetime imaging microscopy, and single- and multi-photon excitation. Edited by pioneers in this field, with contributions from leading experts, the book includes an overview of complementary techniques that help researchers beginning FLIM research. It offers a comprehensive treatment of fundamental principles, instrumentation, analytical methods, and applications. It also provides an overview of the label-free contrast available from lifetime measurements of tissue autofluorescence and the prospects for exploiting this for clinical applications and biomedical research including drug discovery.

Reflecting the expanding field’s need for reliable protocols, Fluorescence Spectroscopy and Microscopy: Methods and Protocols offers techniques from a worldwide team of experts on this versatile and vital subject. The topics covered fall into four broad categories: steady-state fluorescence spectroscopy, time-resolved fluorescence spectroscopy, fluorescent probe development, and the various sub-categories of fluorescence microscopy, such as fluorescence recovery after photobleaching (FRAP), live cell FRET imaging (FRETim), fluorescence lifetime imaging (FLIM), fluorescence fluctuation spectroscopy (FFS), and single-molecule fluorescence spectroscopy (smFS). Written as a part of the popular Methods in Molecular Biology series, chapters include the kind of unambiguous detail and key implementation advice that proves essential for successful results. Comprehensive and practical, Fluorescence Spectroscopy and Microscopy: Methods and Protocols aims to guide both ‘novice’ and established scientists toward furthering their research with these invaluable techniques.

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.

The increased use of fluorescence techniques is greatly enhanced by the improved instrumentation pioneered by inventive scientists and now made available commercially by several high-tech companies. Moreover, the design and development of many new molecular probes with higher selectivity for specific microenvironmental properties has stimulated many new researchers to employ fluorescence techniques for solving their problems. This topic book, the second in his series, reflects this exciting scientific progress and deals, among others, with new approaches and new probes in fluorescence spectroscopy, single molecule fluorescence, applications in biomembrane and enzyme studies and imaging of living cells.

This is a practical introduction to single molecule fluorescence experiments, the analysis of the data, and applications of the techniques to the study of biological structure and function.

This handbook provides a straightforward introduction to spectroscopy, showing what it can do and how it does it, together with a clear, integrated and objective account of the wealth of information that can be derived from spectra. The sequence of chapters covers a wide range of the electromagnetic spectrum, and the physical processes involved, from nuclear phenomena to molecular rotation processes. - A day-by-day laboratory guide: its design based on practical knowledge of spectroscopists at universities, industries and research institutes - A well-structured information source containing methods and applications sections framed by sections on general topics - Guides users to a decision about which spectroscopic method and which instrumentation will be the most appropriate to solve their own practical problem - Rapid access to essential information - Correct analysis of a huge number of measured spectra data and smart use of such information sources as databases and spectra libraries

X-Ray fluorescence analysis is an established technique for non-destructive elemental materials analysis. This book gives a user-oriented practical guidance to the application of this method. The book gives a survey of the theoretical fundamentals, analytical instrumentation, software for data processing, various excitation regimes including gracing incidents and microfocus measurements, quantitative analysis, applications in routine and micro analysis, mineralogy, biology, medicine, criminal investigations, archeology, metallurgy, abrasion, microelectronics, environmental air and water analysis. This book is the bible of X-Ray fluorescence analysis. It gives the basic knowledge on this technique, information on analytical equipment and guides the reader to the various applications. It appeals to researchers, analytically active engineers and advanced students.