Dual-color fluorescence cross-correlation spectroscopy on a Single plane illumination microscope (SPIM-FCCS)
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-correlatio...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
28 Jan 2014
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| In: |
Optics express
Year: 2014, Volume: 22, Issue: 3, Pages: 2358-2375 |
| ISSN: | 1094-4087 |
| DOI: | 10.1364/OE.22.002358 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1364/OE.22.002358 Verlag, lizenzpflichtig, Volltext: https://www.osapublishing.org/oe/abstract.cfm?uri=oe-22-3-2358 
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| Author Notes: | Jan Wolfgang Krieger, Anand Pratap Singh, Christoph S. Garbe, Thorsten Wohland, and Jörg Langowski |
| Summary: | 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. |
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| Item Description: | Gesehen am 06.10.2020 |
| Physical Description: | Online Resource |
| ISSN: | 1094-4087 |
| DOI: | 10.1364/OE.22.002358 |