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Proceedings Paper

Optimal acquisition schemes for superresolution localization microscopy of bleachable fluorophores
Author(s): Alex Small; Edward Shore
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Paper 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.

Paper Details

Date Published: 13 February 2012
PDF: 7 pages
Proc. SPIE 8228, Single Molecule Spectroscopy and Superresolution Imaging V, 82280Q (13 February 2012); doi: 10.1117/12.907406
Show Author Affiliations
Alex Small, California State Polytechnic Univ., Pomona (United States)
Edward Shore, California State Polytechnic Univ., Pomona (United States)

Published in SPIE Proceedings Vol. 8228:
Single Molecule Spectroscopy and Superresolution Imaging V
Jörg Enderlein; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling; Ingo Gregor, Editor(s)

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