A programmable and automated optical electrowetting-on-dielectric (oEWOD) driven platform for massively parallel and sequential processing of single cell assay operations†

IF 6.1 2区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS
Lab on a Chip Pub Date : 2024-07-22 DOI:10.1039/D4LC00245H
Lawrence G. Welch, Jasper Estranero, Panagiotis Tourlomousis, Robert C. R. Wootton, Valentin Radu, Carlos González-Fernández, Tim J. Puchtler, Claire M. Murzeau, Nele M. G. Dieckmann, Aya Shibahara, Brooke W. Longbottom, Clare E. Bryant and Emma L. Talbot
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引用次数: 0

Abstract

Recently, there has been an increasing emphasis on single cell profiling for high-throughput screening workflows in drug discovery and life sciences research. However, the biology underpinning these screens is often complex and is insufficiently addressed by singleplex assay screens. Traditional single cell screening technologies have created powerful sets of ‘omic data that allow users to bioinformatically infer biological function, but have as of yet not empowered direct functional analysis at the level of each individual cell. Consequently, screening campaigns often require multiple secondary screens leading to laborious, time-consuming and expensive workflows in which attrition points may not be queried until late in the process. We describe a platform that harnesses droplet microfluidics and optical electrowetting-on-dielectric (oEWOD) to perform highly-controlled sequential and multiplexed single cell assays in massively parallelised workflows to enable complex cell profiling during screening. Soluble reagents or objects, such as cells or assay beads, are encapsulated into droplets of media in fluorous oil and are actively filtered based on size and optical features ensuring only desirable droplets (e.g. single cell droplets) are retained for analysis, thereby overcoming the Poisson probability distribution. Droplets are stored in an array on a temperature-controlled chip and the history of individual droplets is logged from the point of filter until completion of the workflow. On chip, droplets are subject to an automated and flexible suite of operations including the merging of sample droplets and the fluorescent acquisition of assay readouts to enable complex sequential assay workflows. To demonstrate the broad utility of the platform, we present examples of single-cell functional workflows for various applications such as antibody discovery, infectious disease, and cell and gene therapy.

Abstract Image

Abstract Image

用于大规模并行和顺序处理单细胞检测操作的可编程自动电介质光学电润湿(oEWOD)驱动平台。
最近,在药物发现和生命科学研究的高通量筛选工作流程中,人们越来越重视单细胞分析。然而,这些筛选所依据的生物学原理往往十分复杂,单复式检测筛选无法充分解决这一问题。传统的单细胞筛选技术已经创建了功能强大的 "omic "数据集,使用户能够通过生物信息学方法推断生物功能,但迄今为止还不能在每个单细胞水平上进行直接的功能分析。因此,筛选活动往往需要进行多次二次筛选,导致工作流程费力、费时、费钱,其中的损耗点可能要到后期才能查询到。我们介绍了一种利用液滴微流控技术和电介质光学电润湿(oEWOD)的平台,该平台可在大规模并行化工作流程中执行高度可控的顺序和多重单细胞检测,从而在筛选过程中进行复杂的细胞剖析。可溶性试剂或物体(如细胞或检测珠)被封装成液滴状的介质,并根据尺寸和光学特征进行主动过滤,确保只有理想的液滴(如单细胞液滴)才会被保留下来进行分析,从而克服了泊松概率分布的问题。液滴以阵列形式存储在温控芯片上,并记录单个液滴从过滤到完成工作流程的历史。在芯片上,液滴可进行自动化的灵活操作,包括合并样本液滴和荧光检测读数采集,以实现复杂的连续检测工作流程。为了展示该平台的广泛用途,我们举例说明了抗体发现、传染病、细胞和基因治疗等各种应用中的单细胞功能工作流程。
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来源期刊
Lab on a Chip
Lab on a Chip 工程技术-化学综合
CiteScore
11.10
自引率
8.20%
发文量
434
审稿时长
2.6 months
期刊介绍: Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.
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