Microfluidic platform for rapid multiplexed biological sample handling for molecular interrogation

IF 3.6 3区 化学 Q2 CHEMISTRY, ANALYTICAL
Analyst Pub Date : 2025-04-22 DOI:10.1039/d4an01415d
Seth R. T. Zima, Rithvik V. Turaga, Sofia C. Yeates-Delahoz, Jose M. Ayuso
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Abstract

Cell manufacture and immune cell research often require screening large numbers of analytes like proteins, metabolites, and nucleic acids, which usually entail expensive equipment and labor-intensive assays such as flow cytometry. On the other hand, microfluidic devices can generate numerous readouts with reduced sample volume, however they often require specialized equipment and expertise to fabricate and use. To this end, we have designed a microfluidic device based on a well plate design for multiplexed analysis of biochemical analyte readouts. The device enables simultaneous liquid delivery to multiple individual wells, thus reducing the number of pipetting steps required to perform assays. The capillary valves used to confine liquid within the wells are capable of withstanding freeze-thawing, which enables pre-loading of plates with reagent for storage and later use. We performed a bioluminescence ATP assay to quantify T cell viability and found that the device was able to capture the luminescence signal with a high degree of consistency. Finally, we performed a bioluminescence immunoassay to compare secretion of IFN-γ between naïve and activated CD4+ T cells, thereby highlighting the potential for this technology to measure relevant immune markers.

Abstract Image

用于分子询问的快速多路生物样品处理的微流控平台
细胞制造和免疫细胞研究通常需要筛选大量的分析物,如蛋白质、代谢物和核酸,这通常需要昂贵的设备和劳动密集型的分析,如流式细胞术。另一方面,微流体装置可以在减少样本量的情况下产生大量读数,但是它们通常需要专门的设备和专业知识来制造和使用。为此,我们设计了一种基于孔板设计的微流控装置,用于生化分析物读数的多路分析。该设备可以同时将液体输送到多个单独的井中,从而减少了进行检测所需的移液步骤。用于将液体限制在井内的毛细管阀能够承受冻融,这使得可以预先装载试剂以备储存和以后使用。我们进行了生物发光ATP测定来量化T细胞活力,发现该装置能够以高度一致性捕获发光信号。最后,我们进行了生物发光免疫测定,比较naïve和活化的CD4+ T细胞之间IFN-γ的分泌,从而强调了该技术测量相关免疫标志物的潜力。
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来源期刊
Analyst
Analyst 化学-分析化学
CiteScore
7.80
自引率
4.80%
发文量
636
审稿时长
1.9 months
期刊介绍: "Analyst" journal is the home of premier fundamental discoveries, inventions and applications in the analytical and bioanalytical sciences.
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