A microfluidic flow cytometer enabling absolute quantification of single-cell intracellular proteins
文献类型：期刊论文
| 作者 | Li, Xiufeng1,2; Fan, Beiyuan1,2; Cao, Shanshan3; Chen, Deyong1,2; Zhao, Xiaoting4; Men, Dong3; Yue, Wentao4; Wang, Junbo1,2; Chen, Jian1,2 |
| 刊名 | Lab on a chip
 |
| 出版日期 | 2017-09-21 |
| 卷号 | 17期号:18页码:3129-3137 |
| ISSN号 | 1473-0197 |
| DOI | 10.1039/c7lc00546f |
| 通讯作者 | Men, dong() ; Wang, junbo(jbwang@mail.ie.ac.cn) ; Chen, jian(chenjian@mail.ie.ac.cn) |
| 英文摘要 | Quantification of single-cell proteomics provides key insights into cellular heterogeneity while conventional flow cytometry cannot provide absolute quantification of intracellular proteins of single cells due to the lack of calibration approaches. this paper presents a constriction channel (with a cross sectional area smaller than cells) based microfluidic flow cytometer, capable of collecting copy numbers of specific intracellular proteins. in this platform, single cells stained with fluorescence labelled antibodies were forced to squeeze through the constriction channel with the fluorescence intensities quantified and since cells fully filled the constriction channel during the squeezing process, solutions with fluorescence labelled antibodies were flushed into the constriction channel to obtain calibration curves. by combining raw fluorescence data and calibration curves, absolute quantification of intracellular proteins was realized. as a demonstration, copy numbers of beta-actin of single tumour cells were quantified to be 0.90 +/- 0.30 mu m (a549, ncell = 14228), 2.34 +/- 0.70 mu m (mcf 10a, ncell = 2455), and 0.98 +/- 0.65 mu m (hep g2, ncell = 6945). the travelling time for individual cells was quantified to be roughly 10 ms and thus a throughput of 100 cells per s can be achieved. this microfluidic system can be used to quantify the copy numbers of intracellular proteins in a high-throughput manner, which may function as an enabling technique in the field of single-cell proteomics. |
| WOS关键词 | INTER-LABORATORY VARIATION ; MASS CYTOMETRY ; CANCER-CELLS ; T-CELLS ; SYSTEM ; IDENTIFICATION ; HETEROGENEITY ; ANTIBODIES ; SUSPENSION ; CYTOKINES |
| WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics |
| WOS类目 | Biochemical Research Methods ; Chemistry, Multidisciplinary ; Chemistry, Analytical ; Nanoscience & Nanotechnology |
| 语种 | 英语 |
| WOS记录号 | WOS:000410405600013 |
| 出版者 | ROYAL SOC CHEMISTRY |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2373380 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Men, Dong; Wang, Junbo; Chen, Jian |
| 作者单位 | 1.Chinese Acad Sci, Inst Elect, State Key Lab Transducer Technol, Beijing, Peoples R China 2.Univ Chinese Acad Sci, Beijing, Peoples R China 3.Chinese Acad Sci, Wuhan Inst Virol, Wuhan, Hubei, Peoples R China 4.Capital Med Univ, Beijing Chest Hosp, Dept Cellular & Mol Biol, Beijing, Peoples R China |
| 推荐引用方式 GB/T 7714 | Li, Xiufeng,Fan, Beiyuan,Cao, Shanshan,et al. A microfluidic flow cytometer enabling absolute quantification of single-cell intracellular proteins[J]. Lab on a chip,2017,17(18):3129-3137. |
| APA | Li, Xiufeng.,Fan, Beiyuan.,Cao, Shanshan.,Chen, Deyong.,Zhao, Xiaoting.,...&Chen, Jian.(2017).A microfluidic flow cytometer enabling absolute quantification of single-cell intracellular proteins.Lab on a chip,17(18),3129-3137. |
| MLA | Li, Xiufeng,et al."A microfluidic flow cytometer enabling absolute quantification of single-cell intracellular proteins".Lab on a chip 17.18(2017):3129-3137. |
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来源：武汉病毒研究所
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