Doctoral Program in the Chemical and Biological Sciences and ‡Department of Chemistry, The Scripps Research Institute , 130 Scripps Way, Jupiter, Florida 33458, United States.
Anal Chem. 2017 Dec 19;89(24):13227-13234. doi: 10.1021/acs.analchem.7b03070. Epub 2017 Nov 28.
Automated and reproducible sample handling is a key requirement for high-throughput compound screening and currently demands heavy reliance on expensive robotics in screening centers. Integrated droplet microfluidic screening processors are poised to replace robotic automation by miniaturizing biochemical reactions to the droplet scale. These processors must generate, incubate, and sort droplets for continuous droplet screening, passively handling millions of droplets with complete uniformity, especially during the key step of sample incubation. Here, we disclose an integrated microfluidic emulsion creamer that packs ("creams") assay droplets by draining away excess oil through microfabricated drain channels. The drained oil coflows with creamed emulsion and then reintroduces the oil to disperse the droplets at the circuit terminus for analysis. Creamed emulsion assay incubation time dispersion was 1.7%, 3-fold less than other reported incubators. The integrated, continuous emulsion creamer (ICEcreamer) was used to miniaturize and optimize measurements of various enzymatic activities (phosphodiesterase, kinase, bacterial translation) under multiple- and single-turnover conditions. Combining the ICEcreamer with current integrated microfluidic DNA-encoded library bead processors eliminates potentially cumbersome instrumentation engineering challenges and is compatible with assays of diverse target class activities commonly investigated in drug discovery.
自动化且可重现的样本处理是高通量化合物筛选的关键要求，目前在筛选中心严重依赖昂贵的机器人。集成液滴微流控筛选处理器通过将生化反应微型化到液滴尺度，有望取代机器人自动化。这些处理器必须生成、孵育和分选液滴以进行连续液滴筛选，在样品孵育的关键步骤中，被动地以完全均匀的方式处理数百万个液滴。在这里，我们公开了一种集成的微流控乳液搅拌器，该搅拌器通过微加工排油通道排走多余的油来“搅拌”（cream）检测液滴。排走的油与搅拌好的乳液一起流动，然后在回路末端重新引入油以分散液滴进行分析。搅拌好的乳液孵育时间分散度为 1.7%，比其他报道的孵育器低 3 倍。集成的连续乳液搅拌器（ICEcreamer）用于微型化和优化各种酶活性（磷酸二酯酶、激酶、细菌翻译）的测量，包括多轮和单轮条件下的测量。将 ICEcreamer 与当前的集成微流控 DNA 编码文库珠处理器结合使用，可以消除潜在的繁琐仪器工程挑战，并且与药物发现中常见的各种靶类活性的检测兼容。
