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首页> 外文期刊>Journal of Micromechanics and Microengineering >A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity
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A simple add-on microfluidic appliance for accurately sorting small populations of cells with high fidelity

机译：一种简单的附加微流控设备，可高保真度地准确分选少量细胞

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Current advances in single cell sequencing, gene expression and proteomics require the isolation of single cells, frequently from a very small source population. In this work we describe the design and characterization of a manually operated microfluidic cell sorter that (1) can accurately sort single or small groups of cells from very small cell populations with minimal losses, (2) that is easy to operate and that can be used in any laboratory that has a basic fluorescent microscope and syringe pump, (3) that can be assembled within minutes, (4) that can sort cells in very short time (minutes) with minimum cell stress, (5) that is cheap and reusable. This microfluidic sorter is made from hard plastic material (PMMA) into which microchannels are directly milled with hydraulic diameter of 70 μm. Inlet and outlet reservoirs are drilled through the chip. Sorting occurs through hydrodynamic switching ensuring low hydrodynamic shear stresses, which were modeled and experimentally confirmed to be below the cell damage threshold. Manually operated, the maximum sorting frequencies were approximately 10 cells min~(-1). Experiments verified that cell sorting operations could be achieved in as little as 15 min, including the assembly and testing of the sorter. In only one out of ten sorting experiments the sorted cells were contaminated with another cell type. This microfluidic cell sorter represents an important capability for protocols requiring fast isolation of single cells from small number of rare cell populations.
机译：单细胞测序，基因表达和蛋白质组学的最新进展要求分离单细胞，通常是从非常小的来源群体中分离出来。在这项工作中，我们描述了手动操作的微流细胞分选仪的设计和表征，该分选仪（1）可以从很小的细胞群中准确分选单个或一小组细胞，而损失最小，（2）易于操作并且可以用于具有基本荧光显微镜和注射泵的任何实验室;（3）可以在数分钟内组装完毕;（4）可以在很短的时间内（几分钟）以最小的细胞应力对细胞进行分选;（5）价格便宜且可重用的。这种微流体分选机由硬塑料材料（PMMA）制成，可直接将微通道研磨成水力直径为70μm的微通道。进水口和出水口都通过切屑钻出。通过流体动力转换进行分类，确保低流体动力剪切应力，该应力已建模并通过实验确认低于细胞损伤阈值。手动操作时，最大分选频率约为10细胞min〜（-1）。实验证明，细胞分选操作可以在短短15分钟内完成，包括分选机的组装和测试。在十个分选实验中，只有一个被分选出的细胞被另一种细胞所污染。对于需要从少量稀有细胞群体中快速分离单个细胞的方案而言，这种微流体细胞分选仪具有重要的功能。

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