An expectation-maximization approach to quantifying protein stoichiometry with single-molecule imaging.

ABSTRACT:

Motivation

Single-molecule localization microscopy (SMLM) is a super-resolution technique capable of rendering nanometer scale images of cellular structures. Recently, much effort has gone into developing algorithms for extracting quantitative features from SMLM datasets, such as the abundance and stoichiometry of macromolecular complexes. These algorithms often require knowledge of the complicated photophysical properties of photoswitchable fluorophores.

Results

Here, we develop a calibration-free approach to quantitative SMLM built upon the observation that most photoswitchable fluorophores emit a geometrically distributed number of blinks before photobleaching. From a statistical model of a mixture of monomers, dimers and trimers, the method employs an adapted expectation

SUBMITTER: Boonkird A 

PROVIDER: S-EPMC9710618 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature