title: Dual-color fluorescence cross-correlation spectroscopy on a single plane illumination microscope (SPIM-FCCS).
abstract: Single plane illumination microscopy based fluorescence correlation spectroscopy (SPIM-FCS) is a new method for imaging FCS in 3D samples, providing diffusion coefficients, flow velocities and concentrations in an imaging mode. Here we extend this technique to two-color fluorescence cross-correlation spectroscopy (SPIM-FCCS), which allows to measure molecular interactions in an imaging mode. We present a theoretical framework for SPIM-FCCS fitting models, which is subsequently used to evaluate several test measurements of in-vitro (labeled microspheres, several DNAs and small unilamellar vesicles) and in-vivo samples (dimeric and monomeric dual-color fluorescent proteins, as well as membrane bound proteins). Our method yields the same quantitative results as the well-established confocal FCCS, but in addition provides unmatched statistics and true imaging capabilities.
authors.forename: Jan Wolfgang
authors.initials: JW
authors.lastname: Krieger
authors.forename: Anand Pratap
authors.initials: AP
authors.lastname: Singh
authors.forename: Christoph S
authors.initials: CS
authors.lastname: Garbe
authors.forename: Thorsten
authors.initials: T
authors.lastname: Wohland
authors.forename: Jörg
authors.initials: J
authors.lastname: Langowski
mesh_terms: D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*