Calibration-free counting of low molecular copy numbers in single DNA-PAINT localization clusters.

ABSTRACT: Single-molecule localization microscopy (SMLM) has revolutionized light microscopy by enabling optical resolution down to a few nanometer. Yet, localization precision commonly does not suffice to visually resolve single subunits in molecular assemblies or multimeric complexes. Because each targeted molecule contributes localizations during image acquisition, molecular counting approaches to reveal the target copy numbers within localization clusters have been persistently proposed since the early days of SMLM, most of which rely on preliminary knowledge of the dye photophysics or on a calibration to a reference. Previously, we developed localization-based fluorescence correlation spectroscopy (lbFCS) as an absolute ensemble counting approach for the SMLM-variant DNA-PAINT (points accumulat

SUBMITTER: Stein J 

PROVIDER: S-EPMC9680712 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature