Quantifying Cellular Dynamics by Fluorescence Resonance Energy Transfer (FRET) Microscopy. Issue 1 (1st April 2013)

• Record Type: Journal Article
• Authors: Grecco, Hernán E.; Bastiaens, Philippe I. H.
• Editors: Crawley, Jacqueline N.; Gerfen, Charles R.; Rogawski, Michael A.; Sibley, David R.; Skolnick, Phil; Wray, Susan
• Is Part Of: Current protocols in neuroscience. Issue 63:Issue 1(2013)
• Page Start: 5.22.1
• Page End: 5.22.14
• Publication Date: 2013-04-01
• Subjects: quantification of cellular processes -- fluorescence microscopy -- Förster resonance energy transfer (FRET) -- acceptor photobleaching -- fluorescence lifetime imaging microscopy (FLIM); Neurosciences -- Laboratory manuals; Neurosciences; Cytological Techniques; Histological Techniques; Nervous System -- anatomy & histology; Neurosciences; Laboratory Manual; Laboratory manuals; 612.8
• DOI: 10.1002/0471142301.ns0522s63
• Languages: English
• ISSNs: 1934-8576

Abstract

Abstract: The cell is a spatially organized system whose function emerges from the complex interaction of molecular components. Such local interaction of nanometer‐sized molecules generates patterns that span throughout the cell. Those patterns, in turn, regulate the molecular interactions. Understanding such simultaneous bidirectional causation requires quantifying the spatio‐temporal progression of biochemical reactions in the context of a living cell. Due to its ability to resolve micrometer‐sized structures, biological microscopy has been instrumental to the discovery and understanding of living systems. Functional fluorescence microscopy allows a cellular dynamic topographic map of proteins to be overlaid with topological information on the causality that determines protein state. Here we describe how Förster/fluorescence resonance energy transfer (FRET) can be used to measure the phosphorylation state of proteins in the context of the cell. Curr. Protoc. Neurosci . 63:5.22.1‐5.22.14. © 2013 by John Wiley & Sons, Inc.