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Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level
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 t...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6265747/ https://www.ncbi.nlm.nih.gov/pubmed/30424565 http://dx.doi.org/10.3390/mi9110588 |
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author | Liu, Lixing Fan, Beiyuan Wang, Diancan Li, Xiufeng Song, Yeqing Zhang, Ting Chen, Deyong Wang, Yixiang Wang, Junbo Chen, Jian |
author_facet | Liu, Lixing Fan, Beiyuan Wang, Diancan Li, Xiufeng Song, Yeqing Zhang, Ting Chen, Deyong Wang, Yixiang Wang, Junbo Chen, Jian |
author_sort | Liu, Lixing |
collection | PubMed |
description | 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 × 10(5) and 334.60 nM for Alexa 488, 314.60–2.11 × 10(5) and 153.98 nM for FITC, and 77.03–5.24 × 10(4) 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 × 10(6)/cell (salivary adenoid cystic carcinoma parental cell line (SACC-83), n(cell) = 13,689) vs. 0.90 ± 0.58 × 10(5)/cell (salivary adenoid cystic carcinoma lung metastasis cell line (SACC-LM), n(cell) = 15,341); 0.89 ± 0.69 × 10(6)/cell (oral carcinoma cell line (CAL 27), n(cell) = 7357) vs. 0.93 ± 0.69 × 10(6)/cell (oral carcinoma lymphatic metastasis cell line (CAL 27-LN2), n(cell) = 6276); and 0.86 ± 0.52 × 10(6)/cell (patient I) vs. 0.85 ± 0.58 × 10(6)/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. |
format | Online Article Text |
id | pubmed-6265747 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-62657472018-12-06 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 Micromachines (Basel) Article 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 × 10(5) and 334.60 nM for Alexa 488, 314.60–2.11 × 10(5) and 153.98 nM for FITC, and 77.03–5.24 × 10(4) 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 × 10(6)/cell (salivary adenoid cystic carcinoma parental cell line (SACC-83), n(cell) = 13,689) vs. 0.90 ± 0.58 × 10(5)/cell (salivary adenoid cystic carcinoma lung metastasis cell line (SACC-LM), n(cell) = 15,341); 0.89 ± 0.69 × 10(6)/cell (oral carcinoma cell line (CAL 27), n(cell) = 7357) vs. 0.93 ± 0.69 × 10(6)/cell (oral carcinoma lymphatic metastasis cell line (CAL 27-LN2), n(cell) = 6276); and 0.86 ± 0.52 × 10(6)/cell (patient I) vs. 0.85 ± 0.58 × 10(6)/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. MDPI 2018-11-12 /pmc/articles/PMC6265747/ /pubmed/30424565 http://dx.doi.org/10.3390/mi9110588 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Liu, Lixing Fan, Beiyuan Wang, Diancan Li, Xiufeng Song, Yeqing Zhang, Ting Chen, Deyong Wang, Yixiang Wang, Junbo Chen, Jian Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title | Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title_full | Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title_fullStr | Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title_full_unstemmed | Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title_short | Microfluidic Analyzer Enabling Quantitative Measurements of Specific Intracellular Proteins at the Single-Cell Level |
title_sort | microfluidic analyzer enabling quantitative measurements of specific intracellular proteins at the single-cell level |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6265747/ https://www.ncbi.nlm.nih.gov/pubmed/30424565 http://dx.doi.org/10.3390/mi9110588 |
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