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A three dimensional thermoplastic microfluidic chip for robust cell capture and highresolution imaging

机译:三维热塑性微流控芯片可实现可靠的细胞捕获和高水平分辨率成像

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摘要

We present a low cost microfluidic chip integrating 3D micro-chambers for the capture and the analysis of cells. This device has a simple design and a small footprint. It allows the implementation of standard biological protocols in a chip format with low volume consumption. The manufacturing process relies on hot-embossing of cyclo olefin copolymer, allowing the development of a low cost and robust device. A 3D design of microchannels was used to induce high flow velocity contrasts in the device and provide a selective immobilization. In narrow distribution channels, the liquid velocity induces a shear stress that overcomes adhesion forces and prevents cell immobilization or clogging. In large 3D chambers, the liquid velocity drops down below the threshold for cell attachment. The devices can be operated in a large range of input pressures and can even be handled manually using simple syringe or micropipette. Even at high flow injection rates, the 3D structures protect the captured cell from shear stress. To validate the performances of our device, we implemented immuno-fluorescence labeling and Fluorescence in Situ Hybridization (FISH) analysis on cancer cell lines and on a patient pleural effusion sample. FISH is a Food and Drug Administration approved cancer diagnostic technique that provides quantitative information about gene and chromosome aberration at the single cell level. It is usually considered as a long and fastidious test in medical diagnosis. This process can be easily implanted in our platform, and high resolution fluorescence imaging can be performed with reduced time and computer intensiveness.These results demonstrate the potential of this chip as a low cost, robust, and versatile tooladapted to complex and demanding protocols for medical diagnosis.
机译:我们提出了一种集成了3D微腔室的低成本微流控芯片,用于捕获和分析细胞。该设备设计简单,占地面积小。它允许以芯片格式实现标准的生物协议,并具有较低的体积消耗。制造过程依赖于环烯烃共聚物的热压花,从而允许开发低成本且坚固的装置。微通道的3D设计用于在设备中引起高流速对比并提供选择性固定。在狭窄的分配通道中,液体速度会产生剪切应力,从而克服粘附力并防止细胞固定或阻塞。在大型3D腔室中,液体速度下降到细胞附着阈值以下。该设备可以在很大的输入压力范围内操作,甚至可以使用简单的注射器或微量移液器手动操作。即使在高流量注入速率下,3D结构也可以保护捕获的细胞免受剪切应力。为了验证我们设备的性能,我们在癌细胞系和患者胸腔积液样品上实施了免疫荧光标记和荧光原位杂交(FISH)分析。 FISH是获得美国食品药品监督管理局(FDA)批准的癌症诊断技术,可在单细胞水平上提供有关基因和染色体畸变的定量信息。在医学诊断中,它通常被认为是一项长期而严格的测试。此过程可以轻松地植入我们的平台中,并且可以在减少时间和减少计算机强度的情况下执行高分辨率荧光成像。这些结果证明了该芯片作为低成本,坚固耐用且用途广泛的工具的潜力适应于复杂而苛刻的医学诊断方案。

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