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RESOLFT nanoscopy with photoswitchable organic fluorophores.

ABSTRACT: Far-field optical nanoscopy has been widely used to image small objects with sub-diffraction-limit spatial resolution. Particularly, reversible saturable optical fluorescence transition (RESOLFT) nanoscopy with photoswitchable fluorescent proteins is a powerful method for super-resolution imaging of living cells with low light intensity. Here we demonstrate for the first time the implementation of RESOLFT nanoscopy for a biological system using organic fluorophores, which are smaller in size and easier to be chemically modified. With a covalently-linked dye pair of Cy3 and Alexa647 to label subcellular structures in fixed cells and by optimizing the imaging buffer and optical parameters, our RESOLFT nanoscopy achieved a spatial resolution of ~74?nm in the focal plane. This method provides a powerful alternative for low light intensity RESOLFT nanoscopy, which enables biological imaging with small organic probes at nanoscale resolution.

SUBMITTER: Kwon J

PROVIDER: S-EPMC4671063 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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RESOLFT nanoscopy with photoswitchable organic fluorophores.

Kwon Jiwoong J Hwang Jihee J Park Jaewan J Han Gi Rim GR Han Kyu Young KY Kim Seong Keun SK

Scientific reports 20151207

Far-field optical nanoscopy has been widely used to image small objects with sub-diffraction-limit spatial resolution. Particularly, reversible saturable optical fluorescence transition (RESOLFT) nanoscopy with photoswitchable fluorescent proteins is a powerful method for super-resolution imaging of living cells with low light intensity. Here we demonstrate for the first time the implementation of RESOLFT nanoscopy for a biological system using organic fluorophores, which are smaller in size and ...[more]

PMID: 26639557

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