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HomoFRET Fluorescence Anisotropy Imaging as a Tool to Study Molecular Self‐Assembly in Live Cells
Author(s) -
Chan Fiona T. S.,
Kaminski Clemens F.,
Kaminski Schierle Gabriele S.
Publication year - 2011
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201000833
Subject(s) - förster resonance energy transfer , nanotechnology , context (archaeology) , molecular imaging , fluorescence , live cell imaging , fluorescence anisotropy , chemistry , materials science , physics , biology , cell , optics , biochemistry , paleontology , microbiology and biotechnology , in vivo
Molecular self‐assembly is a defining feature of numerous biological functions and dysfunctions, ranging from basic cell signalling to diseases mediated by protein aggregation. There is current demand for novel experimental methods to study molecular self‐assembly in live cells, and thereby in its physiological context. Förster resonance energy transfer (FRET) between fluorophores of a single type, known as homoFRET, permits noninvasive detection and quantification of molecular clusters in live cells. It can thus provide powerful insights into the molecular physiology of living systems and disease. HomoFRET is detected by measuring the loss of fluorescence anisotropy upon excitation with polarised light. This article reviews recent key developments in homoFRET fluorescence anisotropy imaging for the detection and quantification of molecular self‐assembly reactions in biological systems. A summary is given of the current state‐of‐the‐art and case studies are presented of successful implementations, highlighting technical aspects which have to be mastered to bridge the gap between proof‐of‐concept experiments and biological discoveries.