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美国卫生研究院文献>Biomicrofluidics
>Miniaturized flow cell with pneumatically-actuated vertical nanoconfinement for single-molecule imaging and manipulation
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Miniaturized flow cell with pneumatically-actuated vertical nanoconfinement for single-molecule imaging and manipulation
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 × 25 to 3 × 3 mm2 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.
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机译:凸透镜诱发限制(CLiC)是一种单分子成像技术,它使用可变形的玻璃流动池轻轻捕获,操纵和可视化微结构和纳米结构中的单个分子,从而实现了广泛的应用。在这里,我们将CLiC流通池小型化,从25×25减小到3×3 mm 2 sup>,并引入了气动限制装置。流通池的小型化可将制造产量提高近两个数量级,并且对样品稀少的制药和诊断应用有利,可将内部体积从微升降低到纳升。装置的气动控制减小了限制梯度并改善了机械稳定性,同时保持了低的自发荧光和与油浸物镜的折射率匹配。为了证明我们的“ mini CLiC”系统,我们将DNA限制在50μnm以下的纳米槽中并对其成像,并具有与Odijk限制方案一致的高DNA延伸率。
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