Current microfluidic methods for studying multicell strains (e.g., m-types) with multienvironments (e.g., n-types) require large numbers of inlets/outlets (m*n), a complicated procedure or expensive machinery. Here, we developed a novel two-layer-integrated method to combine different PDMS microchannel layers with different functions into one chip by a PDMS through-hole array, which improved the design of a PDMS-based microfluidic system. Using this method, we succeeded in converting 2 × m × n inlets/outlets into m + n inlets/outlets and reduced the time cost of loading processing (from m × n to m) of the device for studying multicell strains (e.g., m-types) in varied multitemporal environments (i.e., n-types). Using this device, the dynamic behavior of the cell-stress-response proteins was studied when the glucose concentration decreased from 2% to a series of lower concentrations. Our device could also be widely used in high-throughput studies of various stress responses, and the new concept of a multilayer-integrated fabrication method could greatly improve the design of PDMS-based microfluidic systems.

中文翻译：

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一种用于单细胞水平高通量酵母蛋白质组学动力学分析的新型两层集成微流体装置

当前用于研究具有多环境（例如 n 型）的多细胞菌株（例如 m 型）的微流体方法需要大量的入口/出口 (m*n)、复杂的程序或昂贵的机器。在这里，我们开发了一种新的两层集成方法，通过 PDMS 通孔阵列将具有不同功能的不同 PDMS 微通道层组合到一个芯片中，从而改进了基于 PDMS 的微流体系统的设计。使用这种方法，我们成功地将 2 × m × n 个入口/出口转换为 m + n 个入口/出口，并减少了用于研究多细胞菌株（例如，m -types）在不同的多时态环境中（即 n-types）。使用该设备，当葡萄糖浓度从 2% 降低到一系列较低浓度时，研究了细胞应激反应蛋白的动态行为。我们的设备还可以广泛用于各种应力响应的高通量研究，多层集成制造方法的新概念可以大大改善基于 PDMS 的微流体系统的设计。