Miniaturized flow cell with pneumatically-actuated vertical nanoconfinement for single-molecule imaging and manipulation
Miniaturized flow cell with pneumatically-actuated vertical nanoconfinement for single-molecule imaging and manipulation
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DOI：
10.1063/1.5052005
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发表时间：
2018-09-01
期刊：
影响因子：
3.2
通讯作者：
Leslie, Sabrina R.
中科院分区：
文献类型：
--
作者：
Berard, Daniel J.;Leslie, Sabrina R.
凸透镜诱导约束（CLiC）是一种单分子成像技术，它使用可变形的玻璃流池来轻轻地捕获、操纵和可视化微纳米结构中的单分子，从而实现广泛的应用。在这里，我们缩小了CLiC流池，从25 x 25到3 x 3mm(2)，并引入了气动控制的限制。流动池的小型化将制造吞吐量提高了近两个数量级，并且有利于样品珍贵的制药和诊断应用，显着将内部体积从微升降低到纳升。该装置的气动控制减少了限制梯度，提高了机械稳定性，同时保持了与油浸物镜的低自身荧光和折射率匹配。为了演示我们的“迷你CLiC”系统，我们将DNA限制在低于50 nm的纳米沟槽中并对其进行成像，具有与Odijk约束制度一致的高DNA延伸。AIP出版社出版。
Convex Lens-induced Confinement (CLiC) is a single-molecule imaging technique that uses a deformable glass flow cell to gently trap, manipulate, and visualize single molecules within micro-and nano-structures, to enable a wide range of applications. Here, we miniaturize the CLiC flow cell, from 25 x 25 to 3 x 3 mm(2) and introduce pneumatic control of the confinement. Miniaturization of the flow cell improves fabrication throughput by almost two orders of magnitude and, advantageous for pharmaceutical and diagnostic applications where samples are precious, significantly lowers the internal volume from microliters to nanoliters. Pneumatic control of the device reduces the confinement gradient and improves mechanical stability while maintaining low autofluorescence and refractive index-matching with oil-immersion objectives. To demonstrate our "mini CLiC" system, we confine and image DNA in sub-50 nm nanogrooves, with high DNA extension consistent with the Odijk confinement regime. Published by AIP Publishing.