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Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores.

ABSTRACT: Superresolution imaging techniques based on sequential imaging of sparse subsets of single molecules require fluorophores whose emission can be photoactivated or photoswitched. Because typical organic fluorophores can emit significantly more photons than average fluorescent proteins, organic fluorophores have a potential advantage in super-resolution imaging schemes, but targeting to specific cellular proteins must be provided. We report the design and application of HaloTag-based target-specific azido DCDHFs, a class of photoactivatable push-pull fluorogens which produce bright fluorescent labels suitable for single-molecule superresolution imaging in live bacterial and fixed mammalian cells.

SUBMITTER: Lee HL

PROVIDER: S-EPMC2972741 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature

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Superresolution imaging of targeted proteins in fixed and living cells using photoactivatable organic fluorophores.

Lee Hsiao-lu D HL Lord Samuel J SJ Iwanaga Shigeki S Zhan Ke K Xie Hexin H Williams Jarrod C JC Wang Hui H Bowman Grant R GR Goley Erin D ED Shapiro Lucy L Twieg Robert J RJ Rao Jianghong J Moerner W E WE

Journal of the American Chemical Society 20101101 43

Superresolution imaging techniques based on sequential imaging of sparse subsets of single molecules require fluorophores whose emission can be photoactivated or photoswitched. Because typical organic fluorophores can emit significantly more photons than average fluorescent proteins, organic fluorophores have a potential advantage in super-resolution imaging schemes, but targeting to specific cellular proteins must be provided. We report the design and application of HaloTag-based target-specifi ...[more]

PMID: 20936809

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