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Cryogenic Colocalization Microscopy for Nanometer‐Distance Measurements
Author(s) -
Weisenburger Siegfried,
Jing Bo,
Hänni Dominik,
Reymond Luc,
Schuler Benjamin,
Renn Alois,
Sandoghdar Vahid
Publication year - 2014
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201301080
Subject(s) - colocalization , fluorophore , microscopy , nanometre , resolution (logic) , super resolution microscopy , limiting , materials science , microscope , photon , nanotechnology , optics , physics , chemistry , analytical chemistry (journal) , fluorescence , scanning confocal electron microscopy , computer science , mechanical engineering , engineering , artificial intelligence , biology , microbiology and biotechnology , chromatography
The main limiting factor in spatial resolution of localization microscopy is the number of detected photons. Recently we showed that cryogenic measurements improve the photostability of fluorophores, giving access to Angstrom precision in localization of single molecules. Here, we extend this method to colocalize two fluorophores attached to well‐defined positions of a double‐stranded DNA. By measuring the separations of the fluorophore pairs prepared at different design positions, we verify the feasibility of cryogenic distance measurement with sub‐nanometer accuracy. We discuss the important challenges of our method as well as its potential for further improvement and various applications.