Varying label density allows artifact-free analysis of membrane-protein nanoclusters

Florian Baumgart, Andreas Arnold, Konrad Leskovar, Kaj Staszek, Martin Fölser, Julian Weghuber, Hannes Stockinger, Gerhard Schütz

Research output: Contribution to journalArticlepeer-review

61 Citations (Scopus)

Abstract

We present a method to robustly discriminate clustered from randomly distributed molecules detected with techniques based on single-molecule localization microscopy, such as PALM and STORM. The approach is based on deliberate variation of labeling density, such as titration of fluorescent antibody, combined with quantitative cluster analysis, and it thereby circumvents the problem of cluster artifacts generated by overcounting of blinking fluorophores. The method was used to analyze nanocluster formation in resting and activated immune cells.

Original languageEnglish
Pages (from-to)661-664
Number of pages4
JournalNature Methods
Volume13
Issue number8
DOIs
Publication statusPublished - 28 Jul 2016

Keywords

  • Animals
  • Antibodies, Monoclonal/chemistry
  • Artifacts
  • CHO Cells
  • Cell Membrane/metabolism
  • Cluster Analysis
  • Cricetulus
  • Fluorescent Dyes/chemistry
  • Humans
  • Jurkat Cells
  • Light
  • Membrane Proteins/chemistry
  • Microscopy, Fluorescence/methods
  • Nanostructures/chemistry

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