High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events
High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events
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DOI：
10.1364/oe.15.015656
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发表时间：
2007-11-26
期刊：
影响因子：
3.8
通讯作者：
French, P. M. W.
中科院分区：
文献类型：
--
作者：
Grant, D. M.;McGinty, J.;French, P. M. W.
我们提出了一种用于高速活细胞成像的时域光学切片荧光寿命成像（FLIM）显微镜。这个单光子激发系统结合了宽视场平行像素检测和利用旋转尼普科夫圆盘显微镜的共聚焦切片。它可以以每秒10帧（fps）的速度获取活细胞的荧光寿命图像，允许对细胞动力学和蛋白质相互作用进行高速FLIM，具有高通量细胞成像和筛选应用的潜力。我们演示了该FLIM显微镜的应用，以实时监测在使用环糊精胆固醇消耗后细胞膜脂质顺序的变化，并使用FRET激活活细胞中的小gtp酶Ras。(c) 2007年美国光学学会。
We present a time domain optically sectioned fluorescence lifetime imaging (FLIM) microscope developed for high-speed live cell imaging. This single photon excited system combines wide field parallel pixel detection with confocal sectioning utilizing spinning Nipkow disc microscopy. It can acquire fluorescence lifetime images of live cells at up to 10 frames per second (fps), permitting high-speed FLIM of cell dynamics and protein interactions with potential for high throughput cell imaging and screening applications. We demonstrate the application of this FLIM microscope to real-time monitoring of changes in lipid order in cell membranes following cholesterol depletion using cyclodextrin and to the activation of the small GTP-ase Ras in live cells using FRET. (c) 2007 Optical Society of America.