Molecular Imaging: Fret Microscopy and Spectroscopy

Description

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.

Readership:

Biologists, clinical scientists, microscopists, and biomedical science researchers. Course use: basic courses in live cell imaging, advanced undergraduate or graduate courses in microscopy ,and advanced optical techniques.

Contents

1 Robert Kretsinger: Proteins and the Flow of Information in Cellular Function
2 Zygmunt Gryczynski, Ignaci Gryczynski and Joseph Lakowicz: Basics of Fluorescence and FRET
3 C Michael Stanley: An Introducrion to Filters and Mirrors for FRET
4 Fred Schaufele, Ignacio Demarco and Richard N. Day: FRET Imaging in the Wide-field Microscope
5 Horst Wallrabe and Margarida Barroso: Confocal FRET Microscopy: Study of Clustered Distribution of Receptor-ligand Complexes in Endocytic Membranes
6 James D. Mills, James R. Stone, David O. Okonkwo, Ammasi Periasamy and Gregory A. Helm: Multiphoton FRET Microscopy for Protein Localization in Tissue
7 Ye Chen, Masilamani Elangovan, and Ammasi Periasamy:
8 Anne Kenworthy:
9 Sungchul Hohng and Taekjip Ha: Single-Molecule FRET
10 Mary Dickinson and Scott Fraser: FRET Measurements using Multispectral Imaging
11 Glen Redford and Robert M. Clegg: Real-time Fluorescence Lifetime Imaging and FRET using Fast Gated Image Intensifiers
12 V. Krishnan Ramanujan, Jian-Hua Zhang, Victoria. E. Centonze, and Brian Herman: Streak Fluorescence Lifetime Imaging: A Novel Technology for Quantitative FRET Imaging
13 Ye Chen and Ammasi Periasamy: Time-Correlated Single Photon Counting Fluroescence Lifetime Imaging - FRET Microscopy for Protein Localization
14 Mohammed Soutto, Yao Xu, and Carl H. Johnson: Bioluminescence Resonance Energy Transfer: Techniques and Potential
15 Keith Berland: Quantifying Molecular Interactions with Fluorescence Correlation Spectroscopy
16 Elenora Keating, Claire M. Brown and Nils O. Petersen: Mapping Molecular Interactions and Transport in Cell Membranes by Image Correlation