Photoswitching microscopy with standard fluorophores

van de Linde, S. and Kasper, R. and Heilemann, M. and Sauer, M. (2008) Photoswitching microscopy with standard fluorophores. Applied Physics B: Lasers and Optics, 93 (4). pp. 725-731. ISSN 0946-2171 (https://doi.org/10.1007/s00340-008-3250-9)

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Abstract

We introduce far-field subdiffraction-resolution fluorescence imaging based on photoswitching of individual standard fluorophores in air-saturated solution. Here, photoswitching microscopy relies on the light-induced switching of organic fluorophores (ATTO 655 and ATTO 680) into long-lived metastable dark states and spontaneous repopulation of the fluorescent state. In the presence of low concentrations (2-10 mM) of reducing, thiol-containing compounds such as ß-mercaptoethylamine or glutathione, the density of fluorescent molecules can be adjusted to enable multiple localizations of individual fluorophores with an experimental accuracy of ̃20 nm. The method requires wide-field illumination with only a single laser beam for readout and photoswitching and provides superresolution fluorescence images of intracellular structures under live cell compatible conditions.

  • Item type:Article
    ID code:77984
    Dates:
    Date
    Event
    19 October 2008
    Published
    2 October 2008
    Accepted
    Subjects:Science > Physics
    Department:Faculty of Science > Physics
    Depositing user: Pure Administrator
    Date deposited:01 Oct 2021 15:25
    Last modified:21 Nov 2024 02:19
    Related URLs:
    URI:https://strathprints.strath.ac.uk/id/eprint/77984
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