Progress in quantitative single-molecule localization microscopy

H. Deschout, A. Shivanandan, P. Annibale, M. Scarselli, A. Radenovic*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

60 Citations (Scopus)

Abstract

With the advent of single-molecule localization microscopy (SMLM) techniques, intracellular proteins can be imaged at unprecedented resolution with high specificity and contrast. These techniques can lead to a better understanding of cell functioning, as they allow, among other applications, counting the number of molecules of a protein specie in a single cell, studying the heterogeneity in protein spatial organization, and probing the spatial interactions between different protein species. However, the use of these techniques for accurate quantitative measurements requires corrections for multiple inherent sources of error, including: overcounting due to multiple localizations of a single fluorophore (i.e., photoblinking), undercounting caused by incomplete photoconversion, uncertainty in the localization of single molecules, sample drift during the long imaging time, and inaccurate image registration in the case of dual-color imaging. In this paper, we review recent efforts that address some of these sources of error in quantitative SMLM and give examples in the context of photoactivated localization microscopy (PALM).

Original languageEnglish
Pages (from-to)5-17
Number of pages13
JournalHistochemistry and Cell Biology
Volume142
Issue number1
DOIs
Publication statusPublished - Jul 2014

Keywords

  • Cluster analysis
  • Co-localization
  • Fluorescent protein
  • Photoactivated localization microscopy (PALM)
  • Quantitative microscopy
  • Single-molecule counting
  • Single-molecule localization microscopy (SMLM)

Fingerprint

Dive into the research topics of 'Progress in quantitative single-molecule localization microscopy'. Together they form a unique fingerprint.

Cite this