Three dimensional time-gated tracking of non-blinking quantum dots in live cells.
Three dimensional time-gated tracking of non-blinking quantum dots in live cells.
复制标题
活细胞中非闪烁量子点的三维时间选通跟踪。
DOI：
10.1117/12.2082943
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Werner,JamesH
中科院分区：
文献类型：
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作者：
DeVore,MatthewS;Stich,DominikG;Keller,AaronM;Ghosh,Yagnaseni;Goodwin,PeterM;Phipps,MaryE;Stewart,MichaelH;Cleyrat,Cédric;Wilson,BridgetS;Lidke,DianeS;Hollingsworth,JenniferA;Werner,JamesH
单粒子跟踪提供了丰富的生物物理过程的信息，如马达蛋白质的运输和扩散在细胞膜。然而，显微镜平面外的运动或用作标记的荧光探针的闪烁通常将观察时间限制为几秒钟。在这里，我们通过使用新型的不闪烁量子点作为探针并采用定制的3D跟踪显微镜来主动跟踪活细胞中的三维（3D）运动来克服这些限制。通过使用脉冲激发和时间门控检测，在细胞环境中改善信噪比。
Single particle tracking has provided a wealth of information about biophysical processes such as motor protein transport and diffusion in cell membranes. However, motion out of the plane of the microscope or blinking of the fluorescent probe used as a label generally limits observation times to several seconds. Here, we overcome these limitations by using novel non-blinking quantum dots as probes and employing a custom 3D tracking microscope to actively follow motion in three dimensions (3D) in live cells. Signal-to-noise is improved in the cellular milieu through the use of pulsed excitation and time-gated detection.