High-throughput single-cell analysis of low copy number beta-galactosidase by a laboratory-built high-sensitivity flow cytometer

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	High-throughput single-cell analysis of low copy number beta-galactosidase by a laboratory-built high-sensitivity flow cytometer
	Yang, Lingling ; Huang, Tianxun ; Zhu, Shaobin ; Zhou, Yingxing ; Jiang, Yunbin ; Wang, Shuo ; Chen, Yuqing ; Wu, Lina ; Yan, Xiaomei ; Yan XM(颜晓梅)
刊名	http://dx.doi.org/10.1016/j.bios.2013.03.078
	2013
关键词	ESCHERICHIA-COLI GENE-EXPRESSION PROTEIN EXPRESSION CHEMICAL-ANALYSIS FLUORESCENCE NOISE QUANTIFICATION LEVEL ABSOLUTE LIMIT
英文摘要	National Natural Science Foundation of China [21225523, 20975087, 90913015, 21027010]; Program for New Century Excellent Talents in University [NCET-07-0729]; NFFTBS [J1210014]; Single-cell analysis is vital in providing insights into the heterogeneity in molecular content and phenotypic characteristics of complex or clonal cell populations. As many essential proteins and most transcription factors are produced at a low copy number, analytical tools with superior sensitivity to enable the analysis of low abundance proteins in single cells are in high demand. beta-galactosidase (beta-gal) has been the standard cellular reporter for gene expression in both prokaryotic and eukaryotic cells. Here we report the development of a high-throughput method for the single-cell analysis of low copy number beta-gal proteins using a laboratory-built high-sensitivity flow cytometer (HSFCM). Upon fluorescence staining with a fluorogenic substrate, quantitative measurements of the basal and near-basal expression of beta-gal in single Escherichia coli BL21(DE3) cells were demonstrated. Statistical distribution can be determined quickly by analyzing thousands of individual cells in 1-2 min, which reveals the heterogeneous expression pattern that is otherwise masked by the ensemble analysis. Combined with the quantitative fluorometric assay and the rapid bacterial enumeration by HSFCM, the beta-gal expression distribution profile could be converted from arbitrary fluorescence units to protein copy numbers per cell. The sensitivity and speed of the HSFCM offers great capability in quantitative analysis of low abundance proteins in single cells, which would help gaining a deeper insight into the heterogeneity and fundamental biological processes in microbial populations. (C) 2013 Elsevier B.V. All rights reserved.
语种	英语
出版者	ELSEVIER ADVANCED TECHNOLOGY
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/88860]
专题	化学化工－已发表论文
推荐引用方式 GB/T 7714	Yang, Lingling,Huang, Tianxun,Zhu, Shaobin,et al. High-throughput single-cell analysis of low copy number beta-galactosidase by a laboratory-built high-sensitivity flow cytometer[J]. http://dx.doi.org/10.1016/j.bios.2013.03.078,2013.
APA	Yang, Lingling.,Huang, Tianxun.,Zhu, Shaobin.,Zhou, Yingxing.,Jiang, Yunbin.,...&颜晓梅.(2013).High-throughput single-cell analysis of low copy number beta-galactosidase by a laboratory-built high-sensitivity flow cytometer.http://dx.doi.org/10.1016/j.bios.2013.03.078.
MLA	Yang, Lingling,et al."High-throughput single-cell analysis of low copy number beta-galactosidase by a laboratory-built high-sensitivity flow cytometer".http://dx.doi.org/10.1016/j.bios.2013.03.078 (2013).