Development of a microfluidic cell-based biosensor integrating a millisecond chemical pulse generator.
Development of a microfluidic cell-based biosensor integrating a millisecond chemical pulse generator.
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DOI：
10.1016/j.bios.2011.01.013
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发表时间：
2011-04
影响因子：
12.6
通讯作者：
Jian Sun-;Pu Chen;Xiaojun Feng;Wei Du;Bifeng Liu
中科院分区：
文献类型：
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作者：
Jian Sun-;Pu Chen;Xiaojun Feng;Wei Du;Bifeng Liu
基于细胞的生物传感器的使用通常受到激动剂诱导的细胞表面受体脱敏的限制。本文研制了一种基于微流控电池的μ生物传感器，用于检测液体环境中的三磷酸腺苷。它由用于脉动进样的毫秒化学脉冲发生器和单个表达内源性P2Y受体的NIH-3T3细胞组成。使用三磷酸腺苷溶液模拟输入信号，以研究μCBB。通过控制十字形微流控芯片两个出口的负压，基于流体动力门控进样产生了ATP溶液的脉冲。在每50s产生100ms的ATP脉冲的情况下，由此产生的钙尖峰的幅度保持在类似的水平，这表明受体的脱敏作用被最小化。因此，所研制的μCBB可用于检测使用时间延长的脉动样品。进一步测定了μCBB检测三磷酸腺苷的灵敏度，并比较了细胞对毫秒级三磷酸腺苷脉冲和长期刺激的反应。
The use of cell-based biosensors is usually limited by agonist-induced desensitization of cell-surface receptors. In this work, a microfluidic cell-based biosensor (μCBB) was developed for the detection of ATP in liquid environments. It consists of a millisecond chemical pulse generator for sample introduction in a pulsatile manner and a single NIH-3T3 cell expressing endogenous P2Y receptors as the sensing element. ATP solutions were used to simulate input signals for investigating the μCBB. By controlling negative pressures on two outlets of a cross-shaped microfluidic chip, pulses of ATP solutions were generated based on hydrodynamic gated injection. With ATP pulses of 100ms every 50s, the amplitude of the resulting calcium spikes maintained at a similar level, suggesting that the receptor desensitization was minimized. Consequently, the developed μCBB could be used for detecting pulsatile samples with extended use times. The sensitivity of the μCBB for detecting ATP was further determined and the cellular responses to millisecond ATP pulses were investigated in comparison to long-term stimulations.