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A fluorescent microbead-based microfluidic immunoassay chip for immune cell cytokine secretion quantification†
Lab on a Chip ( IF 6.1 ) Pub Date : 2018-01-12 00:00:00 , DOI: 10.1039/c7lc01183k
Xin Cui 1 , Ya Liu , Dinglong Hu , Weiyi Qian , Chung Tin , Dong Sun , Weiqiang Chen , Raymond H W Lam
Affiliation  

Quantitative and dynamic analyses of immune cell secretory cytokines are essential for precise determination and characterization of the “immune phenotype” of patients for clinical diagnosis and treatment of immune-related diseases. Although multiple methods including the enzyme-linked immunosorbent assay (ELISA) have been applied for cytokine detection, such measurements remain very challenging in real-time, high-throughput, and high-sensitivity immune cell analysis. In this paper, we report a highly integrated microfluidic device that allows for on-chip isolation, culture, and stimulation, as well as sensitive and dynamic cytokine profiling of immune cells. Such a microfluidic sensing chip is integrated with cytometric fluorescent microbeads for real-time and multiplexed monitoring of immune cell cytokine secretion dynamics, consuming a relatively small extracted sample volume (160 nl) without interrupting the immune cell culture. Furthermore, it is integrated with a Taylor dispersion-based mixing unit in each detection chamber that shortens the immunoassay period down to less than 30 minutes. We demonstrate the profiling of multiple pro-inflammatory cytokine secretions (e.g. interleukin-6, interleukin-8, and tumor necrosis factors) of human peripheral blood mononuclear cells (PBMCs) with a sensitivity of 20 pg ml−1 and a sample volume of 160 nl per detection. Further applications of this automated, rapid, and high-throughput microfluidic immunophenotyping platform can help unleash the mechanisms of systemic immune responses, and enable efficient assessments of the pathologic immune status for clinical diagnosis and immune therapy.

中文翻译:

基于荧光微珠的微流控免疫分析芯片,用于免疫细胞细胞因子分泌定量†

免疫细胞分泌细胞因子的定量和动态分析对于精确确定和表征患者的“免疫表型”对于免疫相关疾病的临床诊断和治疗至关重要。尽管包括酶联免疫吸附测定(ELISA)在内的多种方法已应用于细胞因子检测,但这种测量在实时、高通量、高灵敏度的免疫细胞分析中仍然非常具有挑战性。在本文中,我们报告了一种高度集成的微流体装置,该装置允许芯片上分离、培养和刺激,以及免疫细胞的敏感和动态细胞因子分析。这种微流控传感芯片与细胞计数荧光微珠集成,用于实时、多重监测免疫细胞细胞因子分泌动态,消耗相对较小的提取样品体积(160 nl),而不中断免疫细胞培养。此外,它在每个检测室中集成了基于泰勒色散的混合单元,将免疫测定时间缩短至不到 30 分钟。我们展示了人外周血单核细胞 (PBMC) 的多种促炎细胞因子分泌物(例如白细胞介素 6、白细胞介素 8 和肿瘤坏死因子)的分析,灵敏度为 20 pg ml -1,样本量为 160每次检测 nl。这种自动化、快速、高通量的微流控免疫表型平台的进一步应用有助于释放全身免疫反应机制,并能够有效评估病理免疫状态,用于临床诊断和免疫治疗。
更新日期:2018-01-12
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