Microfluidic perfusion system for automated delivery of temporal gradients to islets of Langerhans.
Microfluidic perfusion system for automated delivery of temporal gradients to islets of Langerhans.
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DOI：
10.1021/ac802579z
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发表时间：
2009-02-01
影响因子：
7.4
通讯作者：
Roper, Michael G.
中科院分区：
文献类型：
--
作者：
Zhang, Xinyu;Roper, Michael G.
开发了一种微流体灌注系统，用于将刺激波形自动递送到装置内的细胞。3层玻璃/聚合物装置包含两个气动泵、12 cm混合通道和0.2 µL细胞室。通过改变泵的流速比，可以产生一系列输出浓度，同时保持恒定的1.43 ± 0.07 µL/min流速。输出浓度可以随时间变化，产生阶跃梯度和其他波形，例如不同振幅和频率的正弦波和三角波。通过比较输出波形的幅度与理论波形的幅度来分析波形。低于0.0098 Hz的频率时，输出波形与输入波形的差异小于20%。为了减少溶液回流到泵中，对阀门程序的操作顺序以及阀座深度的差异蚀刻进行了修改。这些修改将回流减少到未检测到的程度。在这项工作中应用葡萄糖水平梯度来刺激单个胰岛。3和20 mM之间的葡萄糖梯度引起细胞内[Ca 2 +]的明显和强烈的振荡，表明该系统将在未来的细胞生理学研究中是有用的。
A microfluidic perfusion system was developed for automated delivery of stimulant waveforms to cells within the device. The 3-layer glass/polymer device contained two pneumatic pumps, a 12 cm mixing channel, and a 0.2 µL cell chamber. By altering the flow rate ratio of the pumps, a series of output concentrations could be produced while a constant 1.43 ± 0.07 µL/min flow rate was maintained. The output concentrations could be changed in time producing step gradients and other waveforms, such as sine and triangle waves, at different amplitudes and frequencies. Waveforms were analyzed by comparing the amplitude of output waveforms to the amplitude of theoretical waveforms. Below a frequency of 0.0098 Hz, the output waveforms had less than 20% difference than input waveforms. To reduce backflow of solutions into the pumps, the operational sequence of the valving program was modified as well as differential etching of the valve seat depths. These modifications reduced backflow to the point that it was not detected. Gradients in glucose levels were applied in this work to stimulate single islets of Langerhans. Glucose gradients between 3 and 20 mM brought clear and intense oscillations of intracellular [Ca2+] indicating the system will be useful in future studies of cellular physiology.
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