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HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells

Chan FTS, Kaminski CF, and Kaminski Schierle, "HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells", ChemPhysChem, 12, Issue 3, pages 500–509, February 25, 2011 (2011), DOI:10.1002/cphc.201000833 | pdf 


Abstract

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 on novel experimental methods to study molecular self-assembly in live cells, and thereby in its physiological context. Förster Resonance Energy Transfer (FRET) between a single type of fluorophores, known as homoFRET, permits non-invasive 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 on 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.