Microfluidic Flow Cytometry for Single-Cell Protein Analysis.
Microfluidic Flow Cytometry for Single-Cell Protein Analysis.
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DOI：
10.1007/978-1-4939-2987-0_6
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发表时间：
2015
影响因子：
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通讯作者：
Meiye Wu;A. Singh
中科院分区：
文献类型：
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作者：
Meiye Wu;A. Singh
单细胞蛋白质的流式细胞术 (FC) 检测是一种快速、定量的单细胞蛋白质分析方法。微流体技术的最新进展利用了小型化和自动化，使流式细胞术能够分析细胞表面和细胞内蛋白质的单细胞蛋白质谱。在这里，我们描述了微流控 FC 的方法，以及构建能够进行自动化细胞培养、细胞表面受体免疫染色、细胞内磷蛋白和细胞内细胞因子免疫染色以及使用微流式细胞术进行分析的微流控平台的说明。作为我们的平台和协议的演示，我们详细介绍了使用微流体平台在 LPS 刺激的巨噬细胞中 TLR4 受体激活、ERK1/2 磷酸化和 TNFα 产生的概况。
Flow-cytometric (FC) detection of proteins in single cells is a rapid, quantitative method for single-cell protein analysis. Recent advancements in microfluidic technologies have leveraged miniaturization and automation to adapt flow cytometry for analyzing single cell protein profiles both for cell surface and intracellular proteins. Here, we describe the method for microfluidic FC, along with instructions to build a microfluidic platform capable of automated cell culture, cell surface receptor immunostaining, intracellular phosphoprotein and intracellular cytokine immunostaining, and analysis using micro-flow cytometry. As a demonstration of our platform and protocol, we detail the profiling of TLR4 receptor activation, ERK1/2 phosphorylation, and TNFα production in LPS stimulated macrophages using the microfluidic platform.