Dynamic imaging of homo-FRET in live cells by fluorescence anisotropy microscopy.

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TitleDynamic imaging of homo-FRET in live cells by fluorescence anisotropy microscopy.
Publication TypeJournal Article
Year of Publication2012
AuthorsGhosh S, Saha S, Goswami D, Bilgrami S, Mayor S
JournalMethods Enzymol
Date Published2012
KeywordsAnimals, Cell Membrane, Cell Tracking, Drosophila, Fluorescence Polarization, Fluorescence Resonance Energy Transfer, Green Fluorescent Proteins, Image Processing, Computer-Assisted, Lipid Metabolism, Microscopy, Confocal, Microscopy, Fluorescence

Multiple lipid and protein components of the plasma membrane of a living cell are organized, both compositionally and functionally, at different spatial and temporal scales. For instance, Rab protein domains in membranes the clathrin-coated pit, or the immunological synapse are exquisite examples of functional compartmentalization in cell membranes. These assemblies consist in part of nanoscale complexes of lipids and proteins and are necessary to facilitate some specific sorting and signaling functions. It is evident that cellular functions require a regulated spatiotemporal organization of components at the nanoscale, often comprising of countable number of molecular species. Here, we describe multiple homo-FRET-based imaging methods that provide information about nanoscale interactions between fluorescently tagged molecules in live cells, at optically resolved spatial resolution.

Alternate JournalMeth. Enzymol.
PubMed ID22289460
Grant List / / Wellcome Trust / United Kingdom