Ohsugi Yu, Kinjo Masataka
Hokkaido University, Laboratory of Molecular Cell Dynamics, Faculty of Advanced Life Science, Sapporo 001-0021, Japan.
J Biomed Opt. 2009 Jan-Feb;14(1):014030. doi: 10.1117/1.3080723.
We report simultaneous determination of diffusion coefficients at different points of a cell membrane using a multipoint fluorescence correlation spectroscopy (FCS) system. A system carrying seven detection areas in the evanescent field is achieved by using seven optical fibers on the image plane in the detection port of an objective-type total internal reflection FCS (TIR-FCS) system. Fluctuation of fluorescence intensity is monitored and evaluated using seven photomultiplier tubes (PMTs) and a newly constructed multichannel correlator. We demonstrate simultaneous-multipoint FCS, with a 3-mus time resolution, to investigate heterogeneous structures such as cell membranes and membrane-binding molecular dynamics near glass surfaces in live cells.
我们报告了使用多点荧光相关光谱（FCS）系统同时测定细胞膜不同点的扩散系数。通过在物镜型全内反射FCS（TIR-FCS）系统检测端口的图像平面上使用七根光纤，实现了一个在倏逝场中有七个检测区域的系统。利用七个光电倍增管（PMT）和新构建的多通道相关器监测和评估荧光强度的波动。我们展示了具有3微秒时间分辨率的同时多点FCS，以研究活细胞中细胞膜等异质结构以及玻璃表面附近的膜结合分子动力学。
