A digital microfluidic method for multiplexed cell-based apoptosis assays
A digital microfluidic method for multiplexed cell-based apoptosis assays
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DOI：
10.1039/c2lc20893h
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发表时间：
2012-01-01
期刊：
影响因子：
6.1
通讯作者：
Wheeler, Aaron R.
中科院分区：
文献类型：
--
作者：
Bogojevic, Dario;Chamberlain, M. Dean;Wheeler, Aaron R.
数字微流体（DMF）是一种流体处理技术，其中皮升-微升液滴在电极阵列上进行静电操作，最近在化学和生物学中的应用变得流行。DMF设备是可重构的，没有移动部件，并且与传统的高通量筛选基础设施（例如，例如，在一个实施例中，多孔板读数器）。由于这些和其他原因，数字微流体技术已被吹捧为在多重筛选中应用的潜在有用的新工具。在这里，我们介绍了第一个用于实现并行规模的基于细胞的测定的数字微流体平台。由于细胞凋亡作为抗癌药物发现研究的靶点的流行性，选择半胱天冬酶-3活性的荧光细胞凋亡测定作为模型系统。使用数字微流体方法和常规技术（即，移液、抽吸和96孔板。正如预期的那样，数字微流体方法相对于传统技术的试剂消耗减少了33倍。虽然两种类型的方法使用相同的检测器（台式多孔板读数器），但数字微流体方法生成的数据具有较低的检测限和较大的动态范围，因为相对于多孔板中的移液/抽吸，当暴露于DMF的液滴操作时，凋亡细胞不太可能分层。我们认为，这里描述的技术代表了数字微流体技术发展的一个重要里程碑，它是并行细胞筛选和其他应用的有用工具。
Digital microfluidics (DMF), a fluid-handling technique in which picolitre-microlitre droplets are manipulated electrostatically on an array of electrodes, has recently become popular for applications in chemistry and biology. DMF devices are reconfigurable, have no moving parts, and are compatible with conventional high-throughput screening infrastructure (e. g., multiwell plate readers). For these and other reasons, digital microfluidics has been touted as being a potentially useful new tool for applications in multiplexed screening. Here, we introduce the first digital microfluidic platform used to implement parallel-scale cell-based assays. A fluorogenic apoptosis assay for caspase-3 activity was chosen as a model system because of the popularity of apoptosis as a target for anti-cancer drug discovery research. Dose-response profiles of caspase-3 activity as a function of staurosporine concentration were generated using both the digital microfluidic method and conventional techniques (i.e., pipetting, aspiration, and 96-well plates.) As expected, the digital microfluidic method had a 33-fold reduction in reagent consumption relative to the conventional technique. Although both types of methods used the same detector (a benchtop multiwell plate reader), the data generated by the digital microfluidic method had lower detection limits and greater dynamic range because apoptotic cells were much less likely to de-laminate when exposed to droplet manipulation by DMF relative to pipetting/aspiration in multiwell plates. We propose that the techniques described here represent an important milestone in the development of digital microfluidics as a useful tool for parallel cell-based screening and other applications.