Estimating the dynamic range of quantitative single-molecule localization microscopy.

ABSTRACT: In recent years, there have been significant advances in quantifying molecule copy number and protein stoichiometry with single-molecule localization microscopy (SMLM). However, as the density of fluorophores per diffraction-limited spot increases, distinguishing between detection events from different fluorophores becomes progressively more difficult, affecting the accuracy of such measurements. Although essential to the design of quantitative experiments, the dynamic range of SMLM counting techniques has not yet been studied in detail. Here, we provide a working definition of the dynamic range for quantitative SMLM in terms of the relative number of missed localizations or blinks and explore the photophysical and experimental parameters that affect it. We begin with a simple two-state mo

SUBMITTER: Nino DF 

PROVIDER: S-EPMC8511164 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature