Quantifying molecular colocalization in live cell fluorescence microscopy

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

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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
Volume8
Issue number1-2
DOIs
StatePublished - Jan 1 2015

Fingerprint

Molecular interactions
Fluorophores
Fluorescence microscopy
Fluorescence Microscopy
Labeling
Signal to noise ratio
Microscopic examination
Cells
microscopy
Color
Imaging techniques
fluorescence
Molecules
molecular interactions
cells
marking
signal to noise ratios
Signal-To-Noise Ratio
color
thresholds

Keywords

  • 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)

Cite this

Humpert, F., Yahiatène, I., Lummer, M., Sauer, M., & Huser, T. R. (2015). Quantifying molecular colocalization in live cell fluorescence microscopy. Journal of Biophotonics, 8(1-2), 124-132. https://doi.org/10.1002/jbio.201300146

Quantifying molecular colocalization in live cell fluorescence microscopy. / Humpert, Fabian; Yahiatène, Idir; Lummer, Martina; Sauer, Markus; Huser, Thomas R.

In: Journal of Biophotonics, Vol. 8, No. 1-2, 01.01.2015, p. 124-132.

Research output: Contribution to journalArticle

Humpert, F, Yahiatène, I, Lummer, M, Sauer, M & Huser, TR 2015, 'Quantifying molecular colocalization in live cell fluorescence microscopy', Journal of Biophotonics, vol. 8, no. 1-2, pp. 124-132. https://doi.org/10.1002/jbio.201300146
Humpert, Fabian ; Yahiatène, Idir ; Lummer, Martina ; Sauer, Markus ; Huser, Thomas R. / Quantifying molecular colocalization in live cell fluorescence microscopy. In: Journal of Biophotonics. 2015 ; Vol. 8, No. 1-2. pp. 124-132.
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