A microfluidic platform for trapping, releasing and super-resolution imaging of single cells.
A microfluidic platform for trapping, releasing and super-resolution imaging of single cells.
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DOI：
10.1016/j.snb.2016.03.131
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发表时间：
2016-09
期刊：
影响因子：
--
通讯作者：
Seshia AA
中科院分区：
文献类型：
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作者：
Zhou Y;Basu S;Wohlfahrt KJ;Lee SF;Klenerman D;Laue ED;Seshia AA
高效、可靠、长期捕获单个颗粒和细胞。选择性释放或回收被捕获的颗粒/细胞。近分子分辨率超分辨率成像的稳定平台。利用光活化定位显微镜研究小鼠胚胎干细胞。鉴定200 nm大小的着丝粒，精度<15 nm。一种结合流体动力学捕获和微流控阀技术的多层装置已被开发出来，用于单细胞和颗粒的片上操作和成像。该装置包含具有捕获单个颗粒或细胞的捕获通道的流动层和具有阀门通道的控制层，以选择性地控制捕获和释放过程。使用该装置，粒子和细胞已被成功捕获和释放。该装置能够捕获单个颗粒，捕获效率大于95%，只需控制相应的阀门即可捕获、释放和操纵单个颗粒和细胞。此外，发现捕获和释放过程与细胞等生物样品兼容。我们的设备可以在几分钟内稳定固定大量单细胞，大大简化了单细胞表征的实验设置，并为单分子和超分辨率成像提供了稳定的平台。利用超分辨率光激活定位显微镜（PALM）对小鼠胚胎干细胞（mESCs）进行了概念验证的超分辨率成像实验。细胞和细胞核被稳定捕获并成像。约200 nm大小的着丝粒可以被识别，定位精度<15 nm。
Efficient, reliable and long-term trapping of single particles and cells. Selective releasing or retrieving trapped particles/cells. A stable platform for super-resolution imaging at a near molecular resolution. Study of mouse embryonic stem cells using photoactivated localisation microscopy. Identify centromeres of ∼200 nm size with a precision of <15 nm. A multi-layer device, combining hydrodynamic trapping with microfluidic valving techniques, has been developed for on-chip manipulation and imaging of single cells and particles. Such a device contains a flow layer with trapping channels to capture single particles or cells and a control layer with valve channels to selectively control the trap and release processes. Particles and cells have been successfully trapped and released using the proposed device. The device enables the trapping of single particles with a trapping efficiency of greater than 95%, and allows for single particles and cells to be trapped, released and manipulated by simply controlling corresponding valves. Moreover, the trap and release processes are found to be compatible with biological samples like cells. Our device allows stable immobilisation of large numbers of single cells in a few minutes, significantly easing the experiment setup for single-cell characterisation and offering a stable platform for both single-molecule and super-resolution imaging. Proof-of-concept super- resolution imaging experiments with mouse embryonic stem cells (mESCs) have been conducted by exploiting super-resolution photoactivated localisation microscopy (PALM). Cells and nuclei were stably trapped and imaged. Centromeres of ∼200 nm size could be identified with a localisation precision of <15 nm.