Optimal acquisition schemes for superresolution localization microscopy of bleachable fluorophores

Alex Small; Edward Shore

doi:10.1117/12.907406

13 February 2012 Optimal acquisition schemes for superresolution localization microscopy of bleachable fluorophores

Alex Small, Edward Shore

Author Affiliations +

Proceedings Volume 8228, Single Molecule Spectroscopy and Superresolution Imaging V; 82280Q (2012) https://doi.org/10.1117/12.907406
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

Superresolution localization microscopy (e.g. PALM, STORM) builds images with sub-wavelength resolution by analyzing a series of frames containing sparse, non-overlapping single-fluorophore images. A different set of fluorophores is activated in each frame, and the key parameter controlled by the user is the fraction of fluorophores activated. Using variational techniques, we show that the optimal activation probability, which is the result of a tradeoff between speed and accuracy, depends sensitively on two factors: Whether activation and fluorescent emission are controlled by separate wavelengths or by the same wavelength, and (in the single wavelength case) the detailed kinetics of the bleaching process. Here our approach is extended to the case of multi-fluorophore superresolution techniques, where we show that situations that would be ill-suited to singlefluorophore localization are well-suited to multi-fluorophore localization.

Citation Download Citation

Alex Small and Edward Shore "Optimal acquisition schemes for superresolution localization microscopy of bleachable fluorophores", Proc. SPIE 8228, Single Molecule Spectroscopy and Superresolution Imaging V, 82280Q (13 February 2012); https://doi.org/10.1117/12.907406

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