Molecular orientation affects localization accuracy in superresolution far-field fluorescence microscopy.
basic_science · Level V
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- Record sourced from PubMed, PMID 21133355.
- Also identified by DOI 10.1021/nl103472b.
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Abstract
We investigate the cooperative effect of molecular tilt and defocus on fluorophore localization by centroid calculation in far-field superresolution microscopy based on stochastic single molecule switching. If tilt angle and defocus are unknown, the localization contains systematic errors up to about ±125 nm. When imaging rotation-impaired fluorophores of unknown random orientation, the average localization accuracy in three-dimensional samples is typically limited to about ±32 nm, restricting the attainable resolution accordingly.
Medical subject headings
- Fluorescent Dyes
- Microscopy, Fluorescence