Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection.
Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection.
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DOI：
10.1002/elps.201000209
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发表时间：
2010-09
期刊：
影响因子：
2.9
通讯作者：
Soper, Steven Allan
中科院分区：
文献类型：
--
作者：
Okagbare, Paul I.;Soper, Steven Allan
微流控技术是执行高通量筛选 (HTS) 的可行平台，因为它能够自动化流体处理并在较小的占地面积上生成具有高数量密度的流体网络，适合同时进行许多筛选分析的光学询问。虽然大多数 HTS 活动依赖于荧光，但阅读器通常使用点检测并连续处理测定结果，由于低占空比，显着降低了吞吐量或检测灵敏度。为了应对这一挑战，我们在此提出了将高密度微流体网络封装到大视场 (FoV) 超灵敏荧光检测系统的成像区域中的制造方法。流体通道为 1、5 或 10 μm（宽度）、1 μm（深度），节距为 1-10 μm，每个流体处理器均可单独寻址。流体芯片是使用热压花和热熔粘合的成型工具生产的，以封闭流体通道。 40X 显微镜物镜（数值孔径 = 0.75）产生 200 μm 的 FoV，能够使用当前的流体配置询问约 25 个通道。使用具有大视场的超灵敏荧光检测系统将来自每个流体处理器的荧光信号同时转换到电子倍增电荷耦合器件 (EMCCD) 的活动区域。通过对用作模型筛选测定的酶 APE1 的活性进行高通量监测，证明了这些多通道网络在 HTS 中的实用性。
Microfluidics represents a viable platform for performing High Throughput Screening (HTS) due to its ability to automate fluid handling and generate fluidic networks with high number densities over small footprints appropriate for the simultaneous optical interrogation of many screening assays. While most HTS campaigns depend on fluorescence, readers typically use point detection and serially address the assay results significantly lowering throughput or detection sensitivity due to a low duty cycle. To address this challenge, we present here the fabrication of a high density microfluidic network packed into the imaging area of a large field-of-view (FoV) ultrasensitive fluorescence detection system. The fluidic channels were 1, 5 or 10 μm (width), 1 μm (depth) with a pitch of 1–10 μm and each fluidic processor was individually addressable. The fluidic chip was produced from a molding tool using hot embossing and thermal fusion bonding to enclose the fluidic channels. A 40X microscope objective (numerical aperture = 0.75) created a FoV of 200 μm, providing the ability to interrogate ~25 channels using the current fluidic configuration. An ultrasensitive fluorescence detection system with a large FoV was used to transduce fluorescence signals simultaneously from each fluidic processor onto the active area of an electron multiplying charge-coupled device (EMCCD). The utility of these multichannel networks for HTS was demonstrated by carrying out the high throughput monitoring of the activity of an enzyme, APE1, used as a model screening assay.
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