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

• DOI: 10.1364/oe.18.010627
• Publication Date: 2010-05-06T19:46:32Z
• Authors: Thorsten Wohland; Xianke Shi; Jagadish Sankaran; Ernst H.K. Stelzer
• Keywords: Fluorescence Correlation Spectroscopy

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 high spatial resolution in homo- heterogeneous 3D specimens live zebrafish embryos. Our recorded up to 4096 spectra 56 seconds at laser power 60 microW without damaging embryo. This FCS modality more measurements per time more, less photo-toxic sample than confocal methods. In essence, offers opportunities observe biomolecular interactions quantitatively functions multiplexed manner physiologically relevant