Quantifying molecular colocalization in live cell fluorescence microscopy

Fabian Humpert, Idir Yahiatène, Martina Lummer, Markus Sauer, Thomas R Huser

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


One of the most challenging tasks in microscopy is the quantitative identification and characterization of molecular interactions. In living cells this task is typically performed by fluorescent labeling of the interaction partners with spectrally distinct fluorophores and imaging in different color channels. Current methods for determining colocalization of molecules result in outcomes that can vary greatly depending on signal-to-noise ratios, threshold and background levels, or differences in intensity between channels. Here, we present a novel and quantitative method for determining the degree of colocalization in live-cell fluorescence microscopy images for two and more data channels. Moreover, our method enables the construction of images that directly classify areas of high colocalization. (

Original languageEnglish (US)
Pages (from-to)124-132
Number of pages9
JournalJournal of Biophotonics
Issue number1-2
StatePublished - Jan 1 2015


  • Colocalization
  • Confocal microscopy
  • Correlation-matrix
  • Multicolor
  • Norm

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)
  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)


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