Three-dimensional super-resolution and localization of dense clusters of single molecules

When a single molecule is detected in a wide-field microscope, the image approximates the point spread function of the system. However, as the distribution of molecules becomes denser and their images begin to overlap, existing solutions to determine the number of molecules present and their precise...

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Bibliographic Details
Published in:Scientific reports 2014-06, Vol.4 (1), p.5388-5388, Article 5388
Main Authors: Barsic, Anthony, Grover, Ginni, Piestun, Rafael
Format: Article
Language:English
Subjects:
14
14/63
631/1647/328/2236
639/624/1107/328/2238
639/624/1107/510
Data collection
Humanities and Social Sciences
Localization
multidisciplinary
Science
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Summary:When a single molecule is detected in a wide-field microscope, the image approximates the point spread function of the system. However, as the distribution of molecules becomes denser and their images begin to overlap, existing solutions to determine the number of molecules present and their precise three-dimensional locations can tolerate little to no overlap. We propose a localization scheme that can identify several overlapping molecule images while maintaining high localization precision. A solution to this problem involving matched optical and digital techniques, as here proposed, can substantially increase the allowable labeling density and accelerate the data collection time of single-molecule localization microscopy by more than one order of magnitude.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep05388