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

被引：118

作者：

Wohland, Thorsten ^{[1
,2
]}

Shi, Xianke ^{[1
]}

Sankaran, Jagadish ^{[1
,2
]}

Stelzer, Ernst H. K. ^{[3
]}

机构：

[1] Natl Univ Singapore, Dept Chem, Singapore 117543, Singapore

[2] Natl Univ Singapore, Singapore MIT Alliance, Singapore 117576, Singapore

[3] European Mol Biol Lab, Cell Biol & Biophys Unit, D-69117 Heidelberg, Germany

来源：

OPTICS EXPRESS | 2010年 / 18卷 / 10期

关键词：

INTERNAL-REFLECTION FLUORESCENCE; CROSS-CORRELATION; DIFFUSION; TRANSPORT; ACCURATE; EMBRYOS; CELLS;

D O I：

10.1364/OE.18.010627

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

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 mu W 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. (C) 2010 Optical Society of America

引用

页码：10627 / 10641

页数：15

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