A bistable, multiport valve enables microformulators creating microclinical analyzers that reveal aberrant glutamate metabolism in astrocytes derived from a tuberous sclerosis patient
A bistable, multiport valve enables microformulators creating microclinical analyzers that reveal aberrant glutamate metabolism in astrocytes derived from a tuberous sclerosis patient
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DOI：
10.1016/j.snb.2021.129972
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发表时间：
2021-04-28
影响因子：
8.4
通讯作者：
Wikswo, John P.
中科院分区：
文献类型：
--
作者：
Miller, Dusty R.;Schaffer, David K.;Wikswo, John P.
有需要的阀门和泵，在精密和准确的微尺度操作，在他们的应用是通用的，很容易制造。为此，我们开发了一种新的旋转平面多端口阀来准确选择溶液（污染= 5.22 +/- 0.06 ppb）和一种旋转平面蠕动泵来精确控制流体输送（流量= 2.4 +/- 1.7-890 +/- 77 μ L/min）。阀和泵都以适于单层软光刻制造的平面格式实现。这些平面微流体由一个旋转电机驱动，由定制软件远程控制。这两种设备共同构成了一种创新的微制剂，可用于制备多达五种溶液的精确、高保真混合物（与规定混合物的偏差= +/-|0.02 +/- 0.02| %）。该系统重量不到一公斤，占地面积约为500 cm3，产生的压力为255±47 kPa。然后将该微制剂与电化学传感器结合，创建用于实时检测谷氨酸的微临床分析仪（mu CA）。利用mu CA的腔室作为在线生物反应器，我们比较了从对照（CC-3）和携带致病性TSC2突变的结节性硬化症（TSC）患者的人诱导多能干细胞（hiPSCs）分化而来的人星形胶质细胞的谷氨酸稳态。当谷氨酸刺激时，TSC星形胶质细胞摄取的谷氨酸比对照细胞少。这些数据验证了mu CA的分析能力和微制剂的效用，利用它来评估细胞稳态中与疾病相关的改变。
There is a need for valves and pumps that operate at the microscale with precision and accuracy, are versatile in their application, and are easily fabricated. To that end, we developed a new rotary planar multiport valve to faithfully select solutions (contamination = 5.22 +/- 0.06 ppb) and a rotary planar peristaltic pump to precisely control fluid delivery (flow rate = 2.4 +/- 1.7-890 +/- 77 mu L/min). Both the valve and pump were implemented in a planar format amenable to single-layer soft lithographic fabrication. These planar microfluidics were actuated by a rotary motor controlled remotely by custom software. Together, these two devices constitute an innovative microformulator that was used to prepare precise, high-fidelity mixtures of up to five solutions (deviation from prescribed mixture = +/-|0.02 +/- 0.02| %). This system weighed less than a kilogram, occupied around 500 cm3, and generated pressures of 255 +/- 47 kPa. This microformulator was then combined with an electrochemical sensor creating a microclinical analyzer (mu CA) for detecting glutamate in real time. Using the chamber of the mu CA as an in-line bioreactor, we compared glutamate homeostasis in human astrocytes differentiated from humaninduced pluripotent stem cells (hiPSCs) from a control subject (CC-3) and a Tuberous Sclerosis Complex (TSC) patient carrying a pathogenic TSC2 mutation. When challenged with glutamate, TSC astrocytes took up less glutamate than control cells. These data validate the analytical power of the mu CA and the utility of the microformulator by leveraging it to assess disease-related alterations in cellular homeostasis.