A novel microfluidic flow-cytometry for counting numbers of single-cell β-actins

Nanotechnology and Precision Engineering Pub Date : 2020-09-01 DOI:10.1016/j.npe.2020.06.001

Lixing Liu , Beiyuan Fan , Hongyu Yang , Deyong Chen , Shuang Zhang , Junbo Wang , Jian Chen

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引用次数: 1

Abstract

As a house keeping protein with stable expressions, β-actin is used as a loading control in normalization of western blotting. However, the actual numbers of β-actins at the single-cell level remain elusive. Based on a home-developed flow cytometry, single-cell numbers of β-actin from 8 cell types (subtypes) and 2 tumour patient samples were quantified as 9.62 ± 4.29 × 10⁵ (A549, N_cell = 14,242), 6.46 ± 3.34 × 10⁵ (Hep G2, N_cell = 35,932), 1.58 ± 0.90 × 10⁶ (MCF 10A, N_cell = 16,650), 1.08 ± 0.48 × 10⁶ (HeLa, N_cell = 26,151), 7.60 ± 4.34 × 10⁵ (PC3, N_cell = 11,922), 1.10 ± 0.72 × 10⁶ (SACC-83, N_cell = 13,616), 8.58 ± 4.54 × 10⁵ (CAL 27, N_cell = 7271), 9.00 ± 4.69 × 10⁵ (CAL 27-LN2, N_cell = 6222), 8.26 ± 4.48 × 10⁵ (Oral Tumour Patient I, N_cell = 359), and 8.19 ± 5.12 × 10⁵ (Oral Tumour Patient II, N_cell = 175), and were analyzed by statistical approaches including one-way analysis of variance, neural network based pattern recognition and Bayesian estimation, with varied expressions of β-actins among different cell types located. The dataset reported in this study may serve as a reference in future studies of quantitative protein analysis.

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一种用于计数单细胞β-肌动蛋白的新型微流控流式细胞术

β-肌动蛋白作为一种稳定表达的家养蛋白，在western blotting归一化中被用作负载对照。然而，在单细胞水平上β-肌动蛋白的实际数量仍然难以捉摸。自主开发基于流式细胞术,单细胞的β肌动蛋白从8细胞类型(亚型)和2肿瘤患者样本被量化为9.62±4.29×105 (A549 Ncell = 14242), 6.46±3.34×105(消息灵通的G2, Ncell = 35932), 1.58±0.90×106 (MCF 10, Ncell = 16650), 1.08±0.48×106(海拉,Ncell = 26151), 7.60±4.34×105(生物,Ncell = 11922), 1.10±0.72×106 (sacc - 83, Ncell = 13616), 8.58±4.54×105 (Ncell卡尔27日= 7271),9.00±4.69×105 (CAL 27-LN2 Ncell = 6222),分别为8.26±4.48 × 105(口腔肿瘤患者I, Ncell = 359)和8.19±5.12 × 105(口腔肿瘤患者II, Ncell = 175)，采用单因素方差分析、基于神经网络的模式识别和贝叶斯估计等统计方法进行分析，发现β-肌动蛋白在不同细胞类型中的表达存在差异。本研究的数据集可为今后蛋白质定量分析的研究提供参考。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Nanotechnology and Precision Engineering

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