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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 5, Iss. 9 — Jul. 6, 2010

Single Plane Illumination Fluorescence Correlation Spectroscopy (SPIM-FCS) probes inhomogeneous three-dimensional environments

Thorsten Wohland, Xianke Shi, Jagadish Sankaran, and Ernst H.K. Stelzer  »View Author Affiliations


Optics Express, Vol. 18, Issue 10, pp. 10627-10641 (2010)
http://dx.doi.org/10.1364/OE.18.010627


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Abstract

The life sciences require new highly sensitive imaging tools, which allow the quantitative measurement of molecular parameters within a physiological three-dimensional (3D) environment. Therefore, we combined single plane illumination microscopy (SPIM) with camera based fluorescence correlation spectroscopy (FCS). SPIM-FCS provides contiguous particle number and diffusion coefficient images with a high spatial resolution in homo- and heterogeneous 3D specimens and live zebrafish embryos. Our SPIM-FCS recorded up to 4096 spectra within 56 seconds at a laser power of 60 μW without damaging the embryo. This new FCS modality provides more measurements per time and more, less photo-toxic measurements per sample than confocal based methods. In essence, SPIM-FCS offers new opportunities to observe biomolecular interactions quantitatively and functions in a highly multiplexed manner within a physiologically relevant 3D environment.

© 2010 OSA

OCIS Codes
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(300.6280) Spectroscopy : Spectroscopy, fluorescence and luminescence

ToC Category:
Microscopy

History
Original Manuscript: March 17, 2010
Revised Manuscript: May 2, 2010
Manuscript Accepted: May 4, 2010
Published: May 6, 2010

Virtual Issues
Vol. 5, Iss. 9 Virtual Journal for Biomedical Optics

Citation
Thorsten Wohland, Xianke Shi, Jagadish Sankaran, and Ernst H.K. Stelzer, "Single Plane Illumination Fluorescence Correlation Spectroscopy (SPIM-FCS) probes inhomogeneous three-dimensional environments," Opt. Express 18, 10627-10641 (2010)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-18-10-10627


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