期刊
OPTICS EXPRESS
卷 22, 期 3, 页码 2358-2375出版社
OPTICAL SOC AMER
DOI: 10.1364/OE.22.002358
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- NUS-BW (National University of Singapore/BadenWurttemberg)
- Heidelberg Graduate School of Mathematical and Computational Methods for the Sciences
- National University of Singapore
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. (C) 2014 Optical Society of America
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