Fluorescence polarization filtering for accurate single molecule localization

Single-Molecule Localization Microscopy (SMLM) has become one of the most important methods of super-resolution fluorescence microscopy. It is based on the precise localization of single molecules in wide-field microscopy images. It is well known that the localization accuracy can show a significant...

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Bibliographic Details
Published in:APL photonics 2020-06, Vol.5 (6), p.061302-061302-7
Main Authors: Nevskyi, Oleksii, Tsukanov, Roman, Gregor, Ingo, Karedla, Narain, Enderlein, Jörg
Format: Article
Language:English
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Summary:Single-Molecule Localization Microscopy (SMLM) has become one of the most important methods of super-resolution fluorescence microscopy. It is based on the precise localization of single molecules in wide-field microscopy images. It is well known that the localization accuracy can show a significant bias if the imaged molecules have a fixed orientation and are located either close to an interface or not exactly within the focal plane of the microscope. In this Letter, we propose a simple solution to this problem, which is based on polarization-resolved imaging. This method can be easily implemented into any existing SMLM setup, and we demonstrate its performance by imaging single dye molecules embedded into a polymer film, which fixes their orientation in space.
ISSN:2378-0967
2378-0967
DOI:10.1063/5.0009904