Ultrasensitive Multiparameter Phenotyping of Rare Cells Using an Integrated Digital-Molecular-Counting Microfluidic Well Plate.
Ultrasensitive Multiparameter Phenotyping of Rare Cells Using an Integrated Digital-Molecular-Counting Microfluidic Well Plate.
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DOI：
10.1002/smll.202101743
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发表时间：
2021-08
期刊：
影响因子：
--
通讯作者：
Kurabayashi K
中科院分区：
文献类型：
--
作者：
Su SH;Song Y;Newstead MW;Cai T;Wu M;Stephens A;Singer BH;Kurabayashi K
集成的微流控细胞表型平台为研究由细胞分泌的细胞因子介导的多种炎症疾病提供了一种有前景的手段。然而，先前集成在微流控平台中的免疫传感器缺乏对促炎细胞因子细胞分泌中的小信号进行高精度检测的灵敏度。这一限制禁止研究人员研究低丰度或存在于小群体的细胞分泌细胞因子。在此，作者提出了一个名为“数字表型板（dPP）”的集成平台，该平台将数字免疫传感器集成到带有片上细胞分析室的微流控芯片中，并演示了超灵敏的细胞细胞因子分泌谱测量。集成传感器对小鼠TNF-α的检出限（LOD）小至0.25 pg mL−1。每个芯片上的细胞实验室限制细胞的人口范围从~ 20-600在排列单细胞捕获微孔。总之，dPP的这些微流控特征同时允许对单个细胞进行精确计数和基于图像的细胞术，同时对多个样品在多种刺激物条件下罕见细胞释放的TNF-α进行平行测量。dPP平台广泛适用于要求高精度、高通量的细胞表型表征。本研究开发了一个细胞表型平台，用于单个微流控芯片上的细胞捕获/培养、数字分子计数免疫传感和基于图像的细胞计数。该平台能够超灵敏的细胞因子检测和多种罕见细胞样本的多参数表型。广泛应用于研究表达淀粉样蛋白的阿尔茨海默病模型小鼠和野生型小鼠的初级小胶质细胞功能表型差异（细胞因子分泌和纤维状淀粉样蛋白- β吞噬）。
Integrated microfluidic cellular phenotyping platforms provide a promising means of studying a variety of inflammatory diseases mediated by cell-secreted cytokines. However, immunosensors integrated in previous microfluidic platforms lack the sensitivity to detect small signals in the cellular secretion of pro-inflammatory cytokines with high precision. This limitation prohibits researchers from studying cells secreting cytokines at low abundance or existing at a small population. Herein, the authors present an integrated platform named the “digital Phenoplate (dPP),” which integrates digital immunosensors into a microfluidic chip with on-chip cell assay chambers, and demonstrates ultrasensitive cellular cytokine secretory profile measurement. The integrated sensors yield a limit of detection (LOD) as small as 0.25 pg mL−1 for mouse TNF-α. Each on-chip cell assay chamber confines cells whose population ranges from ~20–600 in arrayed single-cell trapping microwells. Together, these microfluidic features of the dPP simultaneously permit precise counting and image-based cytometry of individual cells while performing parallel measurements of TNF-α released from rare cells under multiple stimulant conditions for multiple samples. The dPP platform is broadly applicable to the characterization of cellular phenotypes demanding high precision and high throughput. This study develops a cellular phenotyping platform for cell trapping/culture, digital molecular-counting immunosensing, and image-based cytometry on a single microfluidic chip. The platform enables ultrasensitive cytokine detection and multiparametric phenotyping of multiple rare cell samples. It is extensively applied on studying primary microglia’s functional phenotypic differences (cytokine secretion and fibrillar amyloid-beta phagocytosis) between amyloid-expressing Alzheimer’s disease model mice and wild-type mice.
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影响因子：
28.2
作者：
Gust J;Hay KA;Hanafi LA;Li D;Myerson D;Gonzalez-Cuyar LF;Yeung C;Liles WC;Wurfel M;Lopez JA;Chen J;Chung D;Harju-Baker S;Özpolat T;Fink KR;Riddell SR;Maloney DG;Turtle CJ
通讯作者：
Turtle CJ
影响因子：
4.7
作者：
Doll DN;Rellick SL;Barr TL;Ren X;Simpkins JW
通讯作者：
Simpkins JW
影响因子：
41.2
作者：
Jain N;Vogel V
通讯作者：
Vogel V
增强内源信号传导下用于分泌蛋白组分析的微流体
DOI：
10.1016/j.bbrc.2018.02.025
发表时间：
2018-03-04
影响因子：
3.1
作者：
Hu, Qianjiang;Luni, Camilla;Elvassore, Nicola
通讯作者：
Elvassore, Nicola
影响因子：
4.6
作者：
Haque A;Gheibi P;Gao Y;Foster E;Son KJ;You J;Stybayeva G;Patel D;Revzin A
通讯作者：
Revzin A