Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level

Liu, Lixing; Fan, Beiyuan; Wang, Diancan; Li, Xiufeng; Song, Yeqing; Zhang, Ting; Chen, Deyong; Wang, Yixiang; Wang, Junbo; Chen, Jian

doi:10.3390/mi9110588

Open AccessArticle

Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level

by

Lixing Liu

^1,2,†

,

Beiyuan Fan

^1,2,†

,

Diancan Wang

^3,†,

Xiufeng Li

^1,2

,

Yeqing Song

³,

Ting Zhang

^1,2,

Deyong Chen

^1,2,

Yixiang Wang

^3,*,

Junbo Wang

^1,2,* and

Jian Chen

^1,2,*

¹

State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, China

²

University of Chinese Academy of Sciences, Beijing 100190, China

³

Peking University School of Stomatology, Beijing 10081, China

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Micromachines 2018, 9(11), 588; https://doi.org/10.3390/mi9110588

Submission received: 12 October 2018 / Revised: 2 November 2018 / Accepted: 8 November 2018 / Published: 12 November 2018

(This article belongs to the Special Issue Microfluidics for Cells and Other Organisms)

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Abstract

This paper presents a microfluidic instrument capable of quantifying single-cell specific intracellular proteins, which are composed of three functioning modules and two software platforms. Under the control of a LabVIEW platform, a pressure module flushed cells stained with fluorescent antibodies through a microfluidic module with fluorescent intensities quantified by a fluorescent module and translated into the numbers of specific intracellular proteins at the single-cell level using a MATLAB platform. Detection ranges and resolutions of the analyzer were characterized as 896.78–6.78 × 10⁵ and 334.60 nM for Alexa 488, 314.60–2.11 × 10⁵ and 153.98 nM for FITC, and 77.03–5.24 × 10⁴ and 37.17 nM for FITC-labelled anti-beta-actin antibodies. As a demonstration, the numbers of single-cell beta-actins of two paired oral tumor cell types and two oral patient samples were quantified as: 1.12 ± 0.77 × 10⁶/cell (salivary adenoid cystic carcinoma parental cell line (SACC-83), n_cell = 13,689) vs. 0.90 ± 0.58 × 10⁵/cell (salivary adenoid cystic carcinoma lung metastasis cell line (SACC-LM), n_cell = 15,341); 0.89 ± 0.69 × 10⁶/cell (oral carcinoma cell line (CAL 27), n_cell = 7357) vs. 0.93 ± 0.69 × 10⁶/cell (oral carcinoma lymphatic metastasis cell line (CAL 27-LN2), n_cell = 6276); and 0.86 ± 0.52 × 10⁶/cell (patient I) vs. 0.85 ± 0.58 × 10⁶/cell (patient II). These results (1) validated the developed analyzer with a throughput of 10 cells/s and a processing capability of ~10,000 cells for each cell type, and (2) revealed that as an internal control in cell analysis, the expressions of beta-actins remained stable in oral tumors with different malignant levels.

Keywords: instrumentation; microfluidic flow cytometry; intracellular proteins; absolute quantification

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MDPI and ACS Style

Liu, L.; Fan, B.; Wang, D.; Li, X.; Song, Y.; Zhang, T.; Chen, D.; Wang, Y.; Wang, J.; Chen, J. Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level. Micromachines 2018, 9, 588. https://doi.org/10.3390/mi9110588

AMA Style

Liu L, Fan B, Wang D, Li X, Song Y, Zhang T, Chen D, Wang Y, Wang J, Chen J. Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level. Micromachines. 2018; 9(11):588. https://doi.org/10.3390/mi9110588

Chicago/Turabian Style

Liu, Lixing, Beiyuan Fan, Diancan Wang, Xiufeng Li, Yeqing Song, Ting Zhang, Deyong Chen, Yixiang Wang, Junbo Wang, and Jian Chen. 2018. "Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level" Micromachines 9, no. 11: 588. https://doi.org/10.3390/mi9110588

APA Style

Liu, L., Fan, B., Wang, D., Li, X., Song, Y., Zhang, T., Chen, D., Wang, Y., Wang, J., & Chen, J. (2018). Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level. Micromachines, 9(11), 588. https://doi.org/10.3390/mi9110588

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Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level

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