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MMaple: a photoconvertible fluorescent protein for use in multiple imaging modalities.


ABSTRACT: Recent advances in fluorescence microscopy have extended the spatial resolution to the nanometer scale. Here, we report an engineered photoconvertible fluorescent protein (pcFP) variant, designated as mMaple, that is suited for use in multiple conventional and super-resolution imaging modalities, specifically, widefield and confocal microscopy, structured illumination microscopy (SIM), and single-molecule localization microscopy. We demonstrate the versatility of mMaple by obtaining super-resolution images of protein organization in Escherichia coli and conventional fluorescence images of mammalian cells. Beneficial features of mMaple include high photostability of the green state when expressed in mammalian cells and high steady state intracellular protein concentration of functional protein when expressed in E. coli. mMaple thus enables both fast live-cell ensemble imaging and high precision single molecule localization for a single pcFP-containing construct.

SUBMITTER: McEvoy AL 

PROVIDER: S-EPMC3519878 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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mMaple: a photoconvertible fluorescent protein for use in multiple imaging modalities.

McEvoy Ann L AL   Hoi Hiofan H   Bates Mark M   Platonova Evgenia E   Cranfill Paula J PJ   Baird Michelle A MA   Davidson Michael W MW   Ewers Helge H   Liphardt Jan J   Campbell Robert E RE  

PloS one 20121211 12


Recent advances in fluorescence microscopy have extended the spatial resolution to the nanometer scale. Here, we report an engineered photoconvertible fluorescent protein (pcFP) variant, designated as mMaple, that is suited for use in multiple conventional and super-resolution imaging modalities, specifically, widefield and confocal microscopy, structured illumination microscopy (SIM), and single-molecule localization microscopy. We demonstrate the versatility of mMaple by obtaining super-resolu  ...[more]

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