Flow characterization and patch clamp dose responses using jet microfluidics in a tubeless microfluidic device.
Flow characterization and patch clamp dose responses using jet microfluidics in a tubeless microfluidic device.
复制标题
在无管微流体装置中使用喷射微流体进行流动表征和膜片钳剂量响应。
DOI：
10.1016/j.jneumeth.2017.08.025
复制
发表时间：
2017
影响因子：
3
通讯作者：
Williams,JustinC
中科院分区：
文献类型：
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作者：
Resto,PedroJ;Bhat,Abhishek;Stava,Eric;Lor,Chong;Merriam,Elliot;Diaz-Rivera,RubenE;Pearce,Robert;Blick,Robert;Williams,JustinC
背景表面张力被动泵送是一种无需泵、管道或阀门即可驱动流动的方法，利用小液滴内的压力通过微流体通道移动液体。这些类型的无管装置通常用于细胞生物学。在此，我们介绍了使用无管装置作为膜片钳电生理学的流体交换平台。新方法利用高速液滴和射流的惯性来产生流动并执行溶液的动态混合。然后将它们流过贴片下 GABA 转染的 HEK 细胞，以进行剂量反应分析。结果使用 TIRF 成像和电记录来研究微流体装置的流体交换特性，导致 0-90% 的流体交换时间为数百毫秒。 COMSOL 用于模拟设备内的流动和流体交换。膜片钳实验表明能够使用高速被动泵送及其衍生物来研究峰值剂量反应，但不能用于研究离子通道动力学。与现有方法相比，我们的系统导致流体交换时间比使用标准 12 桶应用系统时慢，并且不如传统方法稳定，但它提供了一个具有附加功能的新平台。结论表面张力被动泵送和无管装置可以以有限的方式用于电生理学。用户可以获得峰值剂量响应，但当前形式的系统无法足够快地进行流体交换来研究大多数离子通道的动力学。
BackgroundSurface tension passive pumping is a way to actuate flow without the need for pumps, tubing or valves by using the pressure inside small drop to move liquid via a microfluidic channel. These types of tubeless devices have typically been used in cell biology. Herein we present the use of tubeless devices as a fluid exchange platform for patch clamp electrophysiology.New methodInertia from high-speed droplets and jets is used to create flow and perform on-the-fly mixing of solutions. These are then flowed over GABA transfected HEK cells under patch in order to perform a dose response analysis.ResultsTIRF imaging and electrical recordings are used to study the fluid exchange properties of the microfluidic device, resulting in 0–90% fluid exchange times of hundreds of milliseconds. COMSOL is used to model flow and fluid exchange within the device. Patch-clamping experiments show the ability to use high-speed passive pumping and its derivatives for studying peak dose responses, but not for studying ion channel kinetics.Comparison with existing method(s)Our system results in fluid exchange times slower than when using a standard 12-barrel application system and is not as stable as traditional methods, but it offers a new platform with added functionality.ConclusionsSurface tension passive pumping and tubeless devices can be used in a limited fashion for electrophysiology. Users may obtain peak dose responses but the system, in its current form, is not capable of fluid exchange fast enough to study the kinetics of most ion channels.
大鼠树突状细胞充当辅助细胞，控制 T 淋巴细胞有丝分裂反应所需的可溶性因子的产生。
DOI：
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发表时间：
1980
影响因子：
11.1
作者：
W. Klinkert;J. Labadie;J. O'Brien;C. Beyer;W. Bowers
通讯作者：
W. Bowers