Dynamic Microfluidic Cytometry for Single-Cell Cellomics: High-Throughput Probing Single-Cell-Resolution Signaling 机翻标题: 暂无翻译,请尝试点击翻译按钮。

来源
Analytical chemistry
年/卷/期
2019 / 91 / 2
页码
1619-1626
ISSN号
0003-2700
作者单位
Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;Huazhong Univ Sci & Technol, Hubei Bioinformat & Mol Imaging Key Lab, Coll Life Sci & Technol,Dept Biomed Engn,Syst Bio, Key Lab Biomed Photon MOE,Wuhan Natl Lab Optoelec, Wuhan 430074, Hubei, Peoples R China;
作者
Chen, Peng;Yan, Shuangqian;Wang, Jie;Guo, Yiran;Dong, Yue;Feng, Xiaojun;Zeng, Xuemei;Li, Yiwei;Du, Wei;Liu, Bi-Feng;
摘要
Cell signaling is a fast, dynamic, and complex process, which controls a variety of critical physiological functions. Methods to investigate such dynamic information, however, suffer from limited throughput in the single-cell level and a lack of precise fluid manipulation. Herein, we present a new strategy, termed dynamic microfluidic cytometry (DMC), for high-throughput probing of G protein-coupled receptor (GPCR) signaling in single-cell resolution (single-cell cellomics analysis) by creatively applied cyclical cell trapping, stimulating, and releasing automatically. Dose-response curves and half-maximal effective concentration (EC50) values for HeLa cells treated with adenosine triphosphate (ATP), histamine (HA), and acetylcholine chloride (ACH) were successfully obtained in the single-cell level. High-throughput single-cell dynamic signaling was further implemented by sequential or simultaneous stimulation, which revealed that different mechanisms were working in triggering intracellular calcium release. In addition, simultaneous stimulation to two different types of cells, HeLa and NIH-3T3 cells, was also successfully realized, which was crucial for online comparison of dynamic signaling of different types of cells. We believe that the proposed DMC provides a versatile means for high-throughput probing single-cell dynamic signaling, which is potentially useful in chemical biology, cell biology, and pharmacology.
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