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Microfluidic active loading of single cells enables analysis of complex clinical specimens

A fundamental trade-off between flow rate and measurement precision limits performance of many single-cell detection strategies, especially for applications that require biophysical measurements from living cells within complex and low-input samples. To address this, we introduce ‘active loading’, a...

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Autores principales: Calistri, Nicholas L., Kimmerling, Robert J., Malinowski, Seth W., Touat, Mehdi, Stevens, Mark M., Olcum, Selim, Ligon, Keith L., Manalis, Scott R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235965/
https://www.ncbi.nlm.nih.gov/pubmed/30429479
http://dx.doi.org/10.1038/s41467-018-07283-x
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author Calistri, Nicholas L.
Kimmerling, Robert J.
Malinowski, Seth W.
Touat, Mehdi
Stevens, Mark M.
Olcum, Selim
Ligon, Keith L.
Manalis, Scott R.
author_facet Calistri, Nicholas L.
Kimmerling, Robert J.
Malinowski, Seth W.
Touat, Mehdi
Stevens, Mark M.
Olcum, Selim
Ligon, Keith L.
Manalis, Scott R.
author_sort Calistri, Nicholas L.
collection PubMed
description A fundamental trade-off between flow rate and measurement precision limits performance of many single-cell detection strategies, especially for applications that require biophysical measurements from living cells within complex and low-input samples. To address this, we introduce ‘active loading’, an automated, optically-triggered fluidic system that improves measurement throughput and robustness by controlling entry of individual cells into a measurement channel. We apply active loading to samples over a range of concentrations (1–1000 particles μL(−1)), demonstrate that measurement time can be decreased by up to 20-fold, and show theoretically that performance of some types of existing single-cell microfluidic devices can be improved by implementing active loading. Finally, we demonstrate how active loading improves clinical feasibility for acute, single-cell drug sensitivity measurements by deploying it to a preclinical setting where we assess patient samples from normal brain, primary and metastatic brain cancers containing a complex, difficult-to-measure mixture of confounding biological debris.
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spelling pubmed-62359652018-11-16 Microfluidic active loading of single cells enables analysis of complex clinical specimens Calistri, Nicholas L. Kimmerling, Robert J. Malinowski, Seth W. Touat, Mehdi Stevens, Mark M. Olcum, Selim Ligon, Keith L. Manalis, Scott R. Nat Commun Article A fundamental trade-off between flow rate and measurement precision limits performance of many single-cell detection strategies, especially for applications that require biophysical measurements from living cells within complex and low-input samples. To address this, we introduce ‘active loading’, an automated, optically-triggered fluidic system that improves measurement throughput and robustness by controlling entry of individual cells into a measurement channel. We apply active loading to samples over a range of concentrations (1–1000 particles μL(−1)), demonstrate that measurement time can be decreased by up to 20-fold, and show theoretically that performance of some types of existing single-cell microfluidic devices can be improved by implementing active loading. Finally, we demonstrate how active loading improves clinical feasibility for acute, single-cell drug sensitivity measurements by deploying it to a preclinical setting where we assess patient samples from normal brain, primary and metastatic brain cancers containing a complex, difficult-to-measure mixture of confounding biological debris. Nature Publishing Group UK 2018-11-14 /pmc/articles/PMC6235965/ /pubmed/30429479 http://dx.doi.org/10.1038/s41467-018-07283-x Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Calistri, Nicholas L.
Kimmerling, Robert J.
Malinowski, Seth W.
Touat, Mehdi
Stevens, Mark M.
Olcum, Selim
Ligon, Keith L.
Manalis, Scott R.
Microfluidic active loading of single cells enables analysis of complex clinical specimens
title Microfluidic active loading of single cells enables analysis of complex clinical specimens
title_full Microfluidic active loading of single cells enables analysis of complex clinical specimens
title_fullStr Microfluidic active loading of single cells enables analysis of complex clinical specimens
title_full_unstemmed Microfluidic active loading of single cells enables analysis of complex clinical specimens
title_short Microfluidic active loading of single cells enables analysis of complex clinical specimens
title_sort microfluidic active loading of single cells enables analysis of complex clinical specimens
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6235965/
https://www.ncbi.nlm.nih.gov/pubmed/30429479
http://dx.doi.org/10.1038/s41467-018-07283-x
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