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Localization-based super-resolution imaging of cellular structures.

Research paper by Pakorn P Kanchanawong, Clare M CM Waterman

Indexed on: 23 Jul '13Published on: 23 Jul '13Published in: Methods in molecular biology (Clifton, N.J.)



Abstract

Fluorescence microscopy allows direct visualization of fluorescently tagged proteins within cells. However, the spatial resolution of conventional fluorescence microscopes is limited by diffraction to ~250 nm, prompting the development of super-resolution microscopy which offers resolution approaching the scale of single proteins, i.e., ~20 nm. Here, we describe protocols for single molecule localization-based super-resolution imaging, using focal adhesion proteins as an example and employing either photoswitchable fluorophores or photoactivatable fluorescent proteins. These protocols should also be easily adaptable to imaging a broad array of macromolecular assemblies in cells whose components can be fluorescently tagged and assemble into high density structures.