title: Quantifying molecular colocalization in live cell fluorescence microscopy.
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.
authors.affiliation: Department of Physics, University of Bielefeld, Bielefeld, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Humpert
authors.forename: Idir
authors.initials: I
authors.lastname: Yahiatène
authors.forename: Martina
authors.initials: M
authors.lastname: Lummer
authors.forename: Markus
authors.initials: M
authors.lastname: Sauer
authors.forename: Thomas
authors.initials: T
authors.lastname: Huser
mesh_terms: D000818:Animals; D001692:Biological Transport; D019556:COS Cells; D002470:Cell Survival; D002522:Chlorocebus aethiops; D003116:Color; D008856:Microscopy, Fluorescence / Q000379:methods*
keywords: colocalization; confocal microscopy; correlation-matrix; multicolor; norm