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.
Original language | English |
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Pages (from-to) | 725-731 |
Number of pages | 7 |
Journal | Applied Physics B: Lasers and Optics |
Volume | 93 |
Issue number | 4 |
DOIs | |
Publication status | Published - 19 Oct 2008 |
Keywords
- photoswitching
- microscopy
- flurophores
- subdiffraction-resolution
- intracellular structures