Journal
ANNUAL REVIEW OF PHYSICAL CHEMISTRY, VOL 61
Volume 61, Issue -, Pages 345-367Publisher
ANNUAL REVIEWS
DOI: 10.1146/annurev.physchem.012809.103444
Keywords
superresolution microscopy; single molecule; PALM; STORM; FPALM; diffraction limit; photoactivation; photoactivatable fluorescent protein; fluorescence imaging
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Funding
- EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT [ZIAHD001609, ZIAHD008850] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING [ZIAEB000071] Funding Source: NIH RePORTER
- Intramural NIH HHS [Z01 HD001609-15] Funding Source: Medline
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Superresolution imaging is a rapidly emerging new field of microscopy that dramatically improves the spatial resolution of light microscopy by over an order of magnitude (similar to 10-20-nm resolution), allowing biological processes to be described at the molecular scale. Here, we discuss a form of superresolution microscopy based on the controlled activation and sampling of sparse subsets of photoconvertible fluorescent molecules. In this single-molecule-based imaging approach, a wide variety of probes have proved valuable, ranging from genetically encodable photoactivatable fluorescent proteins to photoswitchable cyanine dyes. These have been used in diverse applications of superresolution imaging: from three-dimensional, multicolor molecule localization to tracking of nanometric structures and molecules in living cells. Single-molecule-based superresolution imaging thus offers exciting possibilities for obtaining molecular-scale information on biological events occurring at variable timescales.
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