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The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay

Cytometric studies utilizing flow cytometry or multi-well culture plate fluorometry are often limited by a deficit in temporal resolution and a lack of single cell consideration. Unfortunately, many cellular processes, including signaling, motility, and molecular transport, occur transiently over re...

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Autores principales: Byrd IV, Thomas F., Hoang, Loi T., Kim, Eric G., Pfister, Matthew E., Werner, Erik M., Arndt, Stephen E., Chamberlain, Jeffrey W., Hughey, Jacob J., Nguyen, Bao A., Schneibel, Erik J., Wertz, Laura L., Whitfield, Jonathan S., Wikswo, John P., Seale, Kevin T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4038811/
https://www.ncbi.nlm.nih.gov/pubmed/24873950
http://dx.doi.org/10.1038/srep05117
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author Byrd IV, Thomas F.
Hoang, Loi T.
Kim, Eric G.
Pfister, Matthew E.
Werner, Erik M.
Arndt, Stephen E.
Chamberlain, Jeffrey W.
Hughey, Jacob J.
Nguyen, Bao A.
Schneibel, Erik J.
Wertz, Laura L.
Whitfield, Jonathan S.
Wikswo, John P.
Seale, Kevin T.
author_facet Byrd IV, Thomas F.
Hoang, Loi T.
Kim, Eric G.
Pfister, Matthew E.
Werner, Erik M.
Arndt, Stephen E.
Chamberlain, Jeffrey W.
Hughey, Jacob J.
Nguyen, Bao A.
Schneibel, Erik J.
Wertz, Laura L.
Whitfield, Jonathan S.
Wikswo, John P.
Seale, Kevin T.
author_sort Byrd IV, Thomas F.
collection PubMed
description Cytometric studies utilizing flow cytometry or multi-well culture plate fluorometry are often limited by a deficit in temporal resolution and a lack of single cell consideration. Unfortunately, many cellular processes, including signaling, motility, and molecular transport, occur transiently over relatively short periods of time and at different magnitudes between cells. Here we demonstrate the multitrap nanophysiometer (MTNP), a low-volume microfluidic platform housing an array of cell traps, as an effective tool that can be used to study individual unattached cells over time with precise control over the intercellular microenvironment. We show how the MTNP platform can be used for hematologic cancer cell characterization by measuring single T cell levels of CRAC channel modulation, non-translational motility, and ABC-transporter inhibition via a calcein-AM efflux assay. The transporter data indicate that Jurkat T cells exposed to indomethacin continue to accumulate fluorescent calcein for over 60 minutes after calcein-AM is removed from the extracellular space.
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spelling pubmed-40388112014-05-30 The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay Byrd IV, Thomas F. Hoang, Loi T. Kim, Eric G. Pfister, Matthew E. Werner, Erik M. Arndt, Stephen E. Chamberlain, Jeffrey W. Hughey, Jacob J. Nguyen, Bao A. Schneibel, Erik J. Wertz, Laura L. Whitfield, Jonathan S. Wikswo, John P. Seale, Kevin T. Sci Rep Article Cytometric studies utilizing flow cytometry or multi-well culture plate fluorometry are often limited by a deficit in temporal resolution and a lack of single cell consideration. Unfortunately, many cellular processes, including signaling, motility, and molecular transport, occur transiently over relatively short periods of time and at different magnitudes between cells. Here we demonstrate the multitrap nanophysiometer (MTNP), a low-volume microfluidic platform housing an array of cell traps, as an effective tool that can be used to study individual unattached cells over time with precise control over the intercellular microenvironment. We show how the MTNP platform can be used for hematologic cancer cell characterization by measuring single T cell levels of CRAC channel modulation, non-translational motility, and ABC-transporter inhibition via a calcein-AM efflux assay. The transporter data indicate that Jurkat T cells exposed to indomethacin continue to accumulate fluorescent calcein for over 60 minutes after calcein-AM is removed from the extracellular space. Nature Publishing Group 2014-05-30 /pmc/articles/PMC4038811/ /pubmed/24873950 http://dx.doi.org/10.1038/srep05117 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. The images in this article are included in the article's Creative Commons license, unless indicated otherwise in the image credit; if the image is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the image. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Article
Byrd IV, Thomas F.
Hoang, Loi T.
Kim, Eric G.
Pfister, Matthew E.
Werner, Erik M.
Arndt, Stephen E.
Chamberlain, Jeffrey W.
Hughey, Jacob J.
Nguyen, Bao A.
Schneibel, Erik J.
Wertz, Laura L.
Whitfield, Jonathan S.
Wikswo, John P.
Seale, Kevin T.
The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title_full The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title_fullStr The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title_full_unstemmed The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title_short The microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-AM efflux assay
title_sort microfluidic multitrap nanophysiometer for hematologic cancer cell characterization reveals temporal sensitivity of the calcein-am efflux assay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4038811/
https://www.ncbi.nlm.nih.gov/pubmed/24873950
http://dx.doi.org/10.1038/srep05117
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