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

被引：114
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
收藏
页码：10627 / 10641
页数：15
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