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Confinement and Manipulation of Individual Molecules in Attoliter Volumes

机译:在Attoliter量中对单个分子的限制和操纵

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We report observation of fluorescence from individual rhodamine 6G molecules in streams of charged 1-μm-diameter water droplets. With this approach, illumination volumes comparable to diffraction-limited fluorescence microscopy techniques (≤ 500 aL) are achieved, resulting in similarly high contrast between single-molecule fluorescence signals and nonfluorescent background. However, since the fluorescent molecules are confined to electrically charged droplets, in situ electrodynamic manipulation (e.g., focusing, switching or merging) can be accomplished in a straightforward manner, allowing experimental control over both the delivery of molecules of interest to the observation region and the laser-molecule interaction time. As illustrated by photocount statistics that are independent of molecular diffusion and spatial characteristics of the exciation field, individual rhodamine 6G molecules in 1 -μm droplets are reproducibly delivered to a target a few micrometers in diameter at a rate of between 10 and 100 Hz, with laser beam transit times more than 1 order of magnitude longer than diffusion-limited laser-molecule interaction times in equivalent volumes of free solution.
机译:我们报告观察到从带电的1μm直径的水滴流中若丹明6G分子的荧光。通过这种方法,可以实现与衍射极限荧光显微镜技术(≤500 aL)相当的照明体积,从而在单分子荧光信号和非荧光背景之间产生相似的高对比度。但是,由于荧光分子被限制在带电的液滴中,因此可以以直接的方式完成原位电动操作(例如聚焦,切换或合并),从而允许对目标分子向观察区域的传递和激光分子相互作用的时间。如光计数统计所示,该光谱与分子扩散和激发场的空间特征无关,可将1μm液滴中的若丹明6G分子以10至100 Hz的速率可复制地传递至直径为几微米的目标,在相等体积的自由溶液中,激光束的传输时间比扩散受限的激光分子相互作用时间长1个数量级。

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