A pairwise distance distribution correction (DDC) algorithm to eliminate blinking-caused artifacts in SMLM

Christopher H. Bohrer, Xinxing Yang, Shreyasi Thakur, Xiaoli Weng, Brian Tenner, Ryan McQuillen, Brian Ross, Matthew Wooten, Xin Chen, Jin Zhang, Elijah Roberts, Melike Lakadamyali, Jie Xiao

Research output: Contribution to journalArticlepeer-review

Abstract

Single-molecule localization microscopy (SMLM) relies on the blinking behavior of a fluorophore, which is the stochastic switching between fluorescent and dark states. Blinking creates multiple localizations belonging to the same fluorophore, confounding quantitative analyses and interpretations. Here we present a method, termed distance distribution correction (DDC), to eliminate blinking-caused repeat localizations without any additional calibrations. The approach relies on obtaining the true pairwise distance distribution of different fluorophores naturally from the imaging sequence by using distances between localizations separated by a time much longer than the average fluorescence survival time. We show that, using the true pairwise distribution, we can define and maximize the likelihood, obtaining a set of localizations void of blinking artifacts. DDC results in drastic improvements in obtaining the closest estimate of the true spatial organization and number of fluorescent emitters in a wide range of applications, enabling accurate reconstruction and quantification of SMLM images.

Original languageEnglish (US)
Pages (from-to)669-677
Number of pages9
JournalNature Methods
Volume18
Issue number6
DOIs
StatePublished - Jun 2021

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'A pairwise distance distribution correction (DDC) algorithm to eliminate blinking-caused artifacts in SMLM'. Together they form a unique fingerprint.

Cite this