Molecular Imaging: FRET Microscopy and Spectroscopy

by Ammasi Periasamy (Editor), Richard Day (Editor)

Overview

The detection and measurement of the dynamic interactions of proteins within the living cell are critical to our understanding of cell physiology and pathophysiology. With FRET microscopy and spectroscopy techniques, basic and clinical scientists can make such measurements at very high spatial and temporal resolution. But sources of background information about these tools are very limited, so this book fills an important gap. It covers both the basic concepts and theory behind the various FRET microscopy and spectroscopy techniques, and the practical aspects of using the techniques and analyzing the results. The critical tricks for obtaining a good FRET image and precisely quantitating the signals from living specimens at the nanomolecular level are explained. Valuable information about the preparation of biological samples used for FRET image analysis is also provided. The methods covered include different types of microscopy systems and detectors (wide-field, confocal, multi-photon) as well as specialized techniques such as photobleaching FRET, FLIM-FRET microscopy, spectral imaging FRET, single molecule FRET, and time and image correlation spectroscopy. Starting from the basics, the chapters guide readers through the choice of probes to be used for FRET experiments and the selection of the most suitable experimental approaches to address specific biological questions. Up-to-date, consistently organized, and well-illustrated, this unique book will be welcomed by all researchers who wish to use FRET microscopy and spectroscopy techniques.

Product Details

ISBN-13:	9780080536873
Publisher:	Elsevier Science
Publication date:	04/28/2011
Sold by:	Barnes & Noble
Format:	eBook
Pages:	312
File size:	12 MB
Note:	This product may take a few minutes to download.

About the Author

University of Virginia

University of Virginia

Table of Contents

Proteins and the Flow of Information in Cellular Function; Basics of Fluorescence and FRET; An Introducrion to Filters and Mirrors for FRET; FRET Imaging in the Wide-field Microscope; Confocal FRET Microscopy - Study of Clustered Distribution of Receptor-ligand Complexes in Endocytic Membranes; Multiphoton FRET Microscopy for Protein Localization in Tissue; Single-Molecule FRET; FRET Measurements using Multispectral Imaging; Real-time Fluorescence Lifetime Imaging and FRET using Fast Gated Image Intensifiers; Streak FLIM: A Novel Technology for Quantitative FRET Imaging; Time-Correlated Single Photon Counting (TCSPC) FLIM-FRET Microscopy for Protein Localization;Bioluminescence RET (BRET): Techniques and Potential; Quantifying Molecular Interactions with Fluorescence Correlation Spectrosocpy; Mapping Molecular Interactions and Transport in Cell Membranes by Image Correlation Spectroscopy; Time-Correlated Single Photon Counting (TCSPC) FLIM-FRET Microscopy for Protein Localization

Editorial Reviews

5 Stars! from Doody

Reviewer: Bruce A. Fenderson, PhD (Thomas Jefferson University)
Description: Fluorescence microscopy has provided a wealth of information regarding cell structure and function. Förster resonance energy transfer (FRET) microscopy is a special branch of this field, based on the discovery that direct energy transfer can occur between two different fluorescent probes with overlapping emission and excitation spectra. Energy transfer can only occur over a short distance (1-8 nm). This means that it is possible to capture information regarding the proximity of biomolecules in living cells and study intra- and inter-molecular interactions in real time. This beautiful, hardcover book represents an overview of the basic theory and practical application of FRET microscopy. It includes 16 chapters on topics ranging from "FRET microscopy for protein localization" to "mapping molecular interactions and transport in cell membranes." The primary focus of the book is on biophysics and biochemistry.
Purpose: The book is part of the Methods in Physiology Series of the American Physiological Society. According to the editors, its purpose is to address technological developments in FRET microscopy in a way that will "make these methods of molecular imaging easier to understand for all readers." The goals of FRET microscopy are to "provide high spatial and temporal resolution of signals with the goal of localizing and quantifying protein-protein interactions in live specimens."
Audience: It is written primarily for basic science and clinical researchers (graduate students, postdoctoral fellows, and scientists) interested in the application of fluorescence microscopy to the study of cell biology. The book will be valued by a wide audience of researchers interested in chemistry, biophysics, and biochemistry.
Features: This book provides an exciting up-to-date summary of FRET microscopy, including an overview of the fundamental principles, methods of analysis, choice of filters, use of fluorochromes, and tips on data analysis. The range of techniques is broad, and includes confocal microscopy, bioluminescence FRET, photobleaching FRET, and time and image correlation spectroscopy. Each chapter contains an introduction and an extensive list of references. The figures and diagrams are well done and informative and color is used throughout. The book is carefully edited and a joy to read. The authors are outstanding experts in the field.
Assessment: This is a gold mine of information for those interested in using FRET microscopy to study protein-protein interactions. A major strength of this book is that it is focused and yet comprehensive. Each author delves into a different aspect of the field. The book is clearly written, carefully edited, and engaging. It is assumed that the reader has a general understanding of basic physics and biochemistry. I recommend this book highly for all those interested in the art and science of fluorescence microscopy. It will be a valued resource and practical guide for years to come.

Doody's Review Service

"This is a gold mine of information for those interested in using FRET microscopy to study protein-protein interactions. A major strength of this book is that it is focused and yet comprehensive...The book is clearly written, carefully edited, and engaging...It will be a valued resource and practical guide for years to come...5 Stars! (Highest Rating)" —Doody's

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