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Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification

Functional microbeads have been widely applied in molecular identification and other biochemical applications in the past decade, owing to the compatibility with flow cytometry and the commercially available microbeads for a wide range of molecular identification. Nevertheless, there is still a tech...

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Detalles Bibliográficos
Autores principales: Liu, Ya, Li, Jiyu, Hu, Dinglong, Lam, Josh H. M., Sun, Dong, Pang, Stella W., Lam, Raymond H. W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6086689/
https://www.ncbi.nlm.nih.gov/pubmed/30147817
http://dx.doi.org/10.1063/1.5044449
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author Liu, Ya
Li, Jiyu
Hu, Dinglong
Lam, Josh H. M.
Sun, Dong
Pang, Stella W.
Lam, Raymond H. W.
author_facet Liu, Ya
Li, Jiyu
Hu, Dinglong
Lam, Josh H. M.
Sun, Dong
Pang, Stella W.
Lam, Raymond H. W.
author_sort Liu, Ya
collection PubMed
description Functional microbeads have been widely applied in molecular identification and other biochemical applications in the past decade, owing to the compatibility with flow cytometry and the commercially available microbeads for a wide range of molecular identification. Nevertheless, there is still a technical hurdle caused by the significant sample volume required (∼50 μl), limited molecular detection limit (∼20 pg/ml), complicated liquid/microbead handling procedures, and the long reaction time (>2 h). In this work, we optimize the operation of an automated microbead-based microfluidic device for the reagent mixing and the dynamic cytokine detection. In particular, we adopt fluorescence microscopy for quantification of multiple microbeads in each microchamber instead of flow cytometry for a lower detection limit. The operation parameters are then configured for improved measurement performance. As demonstrated, we consider the cytokine secretion of human macrophage-differentiating lymphocytes stimulated by lipopolysaccharides. We examine requirements on the mixing duration, minimal sample volume, and the image analysis scheme for the smaller biosample volume (<5 μl), the lower cytokine detection limit (∼5 pg/ml), and shorter process time (∼30 min). Importantly, this microfluidic strategy can be further extended in the molecular profiling using other functional microbeads for a broad range of biomedical applications.
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spelling pubmed-60866892018-08-24 Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification Liu, Ya Li, Jiyu Hu, Dinglong Lam, Josh H. M. Sun, Dong Pang, Stella W. Lam, Raymond H. W. Biomicrofluidics Regular Articles Functional microbeads have been widely applied in molecular identification and other biochemical applications in the past decade, owing to the compatibility with flow cytometry and the commercially available microbeads for a wide range of molecular identification. Nevertheless, there is still a technical hurdle caused by the significant sample volume required (∼50 μl), limited molecular detection limit (∼20 pg/ml), complicated liquid/microbead handling procedures, and the long reaction time (>2 h). In this work, we optimize the operation of an automated microbead-based microfluidic device for the reagent mixing and the dynamic cytokine detection. In particular, we adopt fluorescence microscopy for quantification of multiple microbeads in each microchamber instead of flow cytometry for a lower detection limit. The operation parameters are then configured for improved measurement performance. As demonstrated, we consider the cytokine secretion of human macrophage-differentiating lymphocytes stimulated by lipopolysaccharides. We examine requirements on the mixing duration, minimal sample volume, and the image analysis scheme for the smaller biosample volume (<5 μl), the lower cytokine detection limit (∼5 pg/ml), and shorter process time (∼30 min). Importantly, this microfluidic strategy can be further extended in the molecular profiling using other functional microbeads for a broad range of biomedical applications. AIP Publishing LLC 2018-08-10 /pmc/articles/PMC6086689/ /pubmed/30147817 http://dx.doi.org/10.1063/1.5044449 Text en © 2018 Author(s). 1932-1058/2018/12(4)/044112/12 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Regular Articles
Liu, Ya
Li, Jiyu
Hu, Dinglong
Lam, Josh H. M.
Sun, Dong
Pang, Stella W.
Lam, Raymond H. W.
Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title_full Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title_fullStr Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title_full_unstemmed Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title_short Microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
title_sort microfluidic implementation of functional cytometric microbeads for improved multiplexed cytokine quantification
topic Regular Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6086689/
https://www.ncbi.nlm.nih.gov/pubmed/30147817
http://dx.doi.org/10.1063/1.5044449
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