Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.

Cellular mechanical properties can reveal physiologically relevant characteristics in many cell types, and several groups have developed microfluidics-based platforms to perform high-throughput single-cell mechanical testing. However, prior work has performed only limited characterization of these p...

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Autores principales:	Brian Li, Kristen L Cotner, Nathaniel K Liu, Stefan Hinz, Mark A LaBarge, Lydia L Sohn
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/64dee0b7e8cd493da09d22b7c60eb10b
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spelling	oai:doaj.org-article:64dee0b7e8cd493da09d22b7c60eb10b2021-12-02T20:13:31ZEvaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.1932-620310.1371/journal.pone.0258982https://doaj.org/article/64dee0b7e8cd493da09d22b7c60eb10b2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0258982https://doaj.org/toc/1932-6203Cellular mechanical properties can reveal physiologically relevant characteristics in many cell types, and several groups have developed microfluidics-based platforms to perform high-throughput single-cell mechanical testing. However, prior work has performed only limited characterization of these platforms' technical variability and reproducibility. Here, we evaluate the repeatability performance of mechano-node-pore sensing, a single-cell mechanical phenotyping platform developed by our research group. We measured the degree to which device-to-device variability and semi-manual data processing affected this platform's measurements of single-cell mechanical properties. We demonstrated high repeatability across the entire technology pipeline even for novice users. We then compared results from identical mechano-node-pore sensing experiments performed by researchers in two different laboratories with different analytical instruments, demonstrating that the mechanical testing results from these two locations are in agreement. Our findings quantify the expectation of technical variability in mechano-node-pore sensing even in minimally experienced hands. Most importantly, we find that the repeatability performance we measured is fully sufficient for interpreting biologically relevant single-cell mechanical measurements with high confidence.Brian LiKristen L CotnerNathaniel K LiuStefan HinzMark A LaBargeLydia L SohnPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10, p e0258982 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Brian Li Kristen L Cotner Nathaniel K Liu Stefan Hinz Mark A LaBarge Lydia L Sohn Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
description	Cellular mechanical properties can reveal physiologically relevant characteristics in many cell types, and several groups have developed microfluidics-based platforms to perform high-throughput single-cell mechanical testing. However, prior work has performed only limited characterization of these platforms' technical variability and reproducibility. Here, we evaluate the repeatability performance of mechano-node-pore sensing, a single-cell mechanical phenotyping platform developed by our research group. We measured the degree to which device-to-device variability and semi-manual data processing affected this platform's measurements of single-cell mechanical properties. We demonstrated high repeatability across the entire technology pipeline even for novice users. We then compared results from identical mechano-node-pore sensing experiments performed by researchers in two different laboratories with different analytical instruments, demonstrating that the mechanical testing results from these two locations are in agreement. Our findings quantify the expectation of technical variability in mechano-node-pore sensing even in minimally experienced hands. Most importantly, we find that the repeatability performance we measured is fully sufficient for interpreting biologically relevant single-cell mechanical measurements with high confidence.
format	article
author	Brian Li Kristen L Cotner Nathaniel K Liu Stefan Hinz Mark A LaBarge Lydia L Sohn
author_facet	Brian Li Kristen L Cotner Nathaniel K Liu Stefan Hinz Mark A LaBarge Lydia L Sohn
author_sort	Brian Li
title	Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
title_short	Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
title_full	Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
title_fullStr	Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
title_full_unstemmed	Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
title_sort	evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/64dee0b7e8cd493da09d22b7c60eb10b
work_keys_str_mv	AT brianli evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping AT kristenlcotner evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping AT nathanielkliu evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping AT stefanhinz evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping AT markalabarge evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping AT lydialsohn evaluatingsourcesoftechnicalvariabilityinthemechanonodeporesensingpipelineandtheireffectonthereproducibilityofsinglecellmechanicalphenotyping
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Evaluating sources of technical variability in the mechano-node-pore sensing pipeline and their effect on the reproducibility of single-cell mechanical phenotyping.

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