Fluorescence polarization filtering for accurate single molecule localization

• Main Authors: Oleksii Nevskyi, Roman Tsukanov, Ingo Gregor, Narain Karedla, Jörg Enderlein
• Format: Article
• Language: English
• Published: AIP Publishing LLC 2020-06-01
• Series: APL Photonics
• Online Access: http://dx.doi.org/10.1063/5.0009904

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.