Continuous operation of microfabricated electrophoresis devices for 24 hours and application to chemical monitoring of living cells.
Continuous operation of microfabricated electrophoresis devices for 24 hours and application to chemical monitoring of living cells.
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DOI：
10.1021/ac901114k
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发表时间：
2009-08-15
影响因子：
7.4
通讯作者：
Kennedy, Robert T.
中科院分区：
文献类型：
--
作者：
Reid, Kendra R.;Kennedy, Robert T.
微芯片电泳是一种新兴的分析技术，具有几个有用的属性，包括快速分离时间，小样品要求和自动化。在许多潜在的应用中，例如化学监测或高通量筛选，可能需要在没有操作员干预的情况下使用系统进行许多分析；然而，微芯片电泳系统的长期运行却很少受到重视。我们开发了一种微芯片电泳系统，该系统可以在24小时内以6秒的间隔自动注射样品，一次可收集14,400次分析。由于芯片上的试剂和电泳缓冲液的降解，该装置原型的连续操作时间限制在2小时；然而，经过修改，使所有试剂连续灌注到设备上的储液池中，确保了始终使用新鲜试剂进行分析，并延长了操作时间。电泳芯片包括细胞灌注室和试剂添加通道，允许通过串行电泳免疫测定对芯片上培养的细胞周围的液体进行化学监测。免疫分析法对胰岛素的检出限为0.4 nM，在整个24小时内产生约4%的相对标准偏差，没有信号漂移的证据。该联合系统用于监测朗格汉斯单个胰岛的胰岛素分泌6至39小时。监测实验表明，胰岛具有分泌动力学，包括长时间非振荡周期后的自发振荡和可能的超昼夜节律。
Microchip electrophoresis is an emerging analytical technology with several useful attributes including rapid separation time, small sample requirements, and automation. In numerous potential applications, such as chemical monitoring or high-throughput screening, it may be desirable to use a system for many analyses without operator intervention; however, long term operation of microchip electrophoresis systems has received little attention. We have developed a microchip electrophoresis system that can automatically inject samples at 6 s intervals for 24 h resulting in collection of 14,400 assays in one session. Continuous operation time of a prototype of the device was limited to 2 h due to degradation of reagents and electrophoresis buffers on the chip; however, modification so that all reagents were continuously perfused into reservoirs on the device ensured fresh reagents were always used for analysis and enabled extended operating sessions. The electrophoresis chip incorporated a cell perfusion chamber and reagent addition channels to allow chemical monitoring of fluid around cells cultured on the chip by serial electrophoretic immunoassays. The immunoassay had detection limits of 0.4 nM for insulin and generated ∼4% relative standard deviation over an entire 24 h period with no evidence of signal drift. The combined system was used to monitor insulin secretion from single islets of Langerhans for 6 to 39 h. The monitoring experiments revealed that islets have secretion dynamics that include spontaneous oscillations after extended non-oscillating periods and possible ultradian rhythms.
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影响因子：
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作者：
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通讯作者：
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Dishinger, John F.;Reid, Kendra R.;Kennedy, Robert T.
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