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Spatially-resolved profiling of carbon nanotube uptake across cell lines

H. D. Summers, P. Rees, J. T-W. Wang, K. T. Al-Jamal, Paul Rees Orcid Logo, Huw Summers Orcid Logo

Nanoscale, Volume: 9, Issue: 20, Pages: 6800 - 6807

Swansea University Authors: Paul Rees Orcid Logo, Huw Summers Orcid Logo

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DOI (Published version): 10.1039/c7nr01561e

Abstract

The internalisation and intra-cellular distribution of carbon nanotubes (CNT) has been quantitatively assessed using imaging flow cytometry. Spatial analysis of the bright field images indicates the presence of a small sub-population (5% of cells) in which the internalised CNTs are packed into prono...

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Published in: Nanoscale
ISSN: 2040-3364 2040-3372
Published: 2017
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URI: https://cronfa.swan.ac.uk/Record/cronfa34685
first_indexed 2017-07-18T20:00:35Z
last_indexed 2020-05-28T18:47:01Z
id cronfa34685
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spelling 2020-05-28T15:54:08.5999857 v2 34685 2017-07-18 Spatially-resolved profiling of carbon nanotube uptake across cell lines 537a2fe031a796a3bde99679ee8c24f5 0000-0002-7715-6914 Paul Rees Paul Rees true false a61c15e220837ebfa52648c143769427 0000-0002-0898-5612 Huw Summers Huw Summers true false 2017-07-18 EAAS The internalisation and intra-cellular distribution of carbon nanotubes (CNT) has been quantitatively assessed using imaging flow cytometry. Spatial analysis of the bright field images indicates the presence of a small sub-population (5% of cells) in which the internalised CNTs are packed into pronounced clusters, visible as dark spots due to strong optical scattering by the nanotubes. The area of these spots can be used as a label-free metric of CNT dose and we assess the relative uptake of charge-neutral CNTs, over a 24 hours exposure period across four cell types: J774 mouse macrophage cells, A549 and Calu-6 human lung cancer cells, and MCF-7 human breast cells. The relative dose as indicated by the spot-area metric closely correlates to results using the same CNT preparation, conjugated to a FITC-label and shows pronounced uptake by the J774 cells leading to a mean dose that is >60% higher than for the other cell types. Spatial evaluation of dosing clusters is also used to quantify differences in uptake by J774 cells of CNTs with different surface functionalisation. While the percentage of CNT-cluster positive cells increases from 5% to 19% when switching from charge-neutral CNTs to poly-cationic, dendron functionalised CNTs, the single cell level analysis of internalised clusters indicates a lower dose per cell of poly-cationic CNTs relative to the charge-neutral CNTs. We concluded that there is dose homeostasis i.e., the population-averaged cellular dose of CNTs remained unchanged. Journal Article Nanoscale 9 20 6800 6807 2040-3364 2040-3372 31 12 2017 2017-12-31 10.1039/c7nr01561e COLLEGE NANME Engineering and Applied Sciences School COLLEGE CODE EAAS Swansea University 2020-05-28T15:54:08.5999857 2017-07-18T15:25:29.5092192 Faculty of Science and Engineering School of Engineering and Applied Sciences - Biomedical Engineering H. D. Summers 1 P. Rees 2 J. T-W. Wang 3 K. T. Al-Jamal 4 Paul Rees 0000-0002-7715-6914 5 Huw Summers 0000-0002-0898-5612 6 0034685-20072017091324.pdf summers2017.pdf 2017-07-20T09:13:24.2670000 Output 2240221 application/pdf Accepted Manuscript true 2018-05-03T00:00:00.0000000 false eng
title Spatially-resolved profiling of carbon nanotube uptake across cell lines
spellingShingle Spatially-resolved profiling of carbon nanotube uptake across cell lines
Paul Rees
Huw Summers
title_short Spatially-resolved profiling of carbon nanotube uptake across cell lines
title_full Spatially-resolved profiling of carbon nanotube uptake across cell lines
title_fullStr Spatially-resolved profiling of carbon nanotube uptake across cell lines
title_full_unstemmed Spatially-resolved profiling of carbon nanotube uptake across cell lines
title_sort Spatially-resolved profiling of carbon nanotube uptake across cell lines
author_id_str_mv 537a2fe031a796a3bde99679ee8c24f5
a61c15e220837ebfa52648c143769427
author_id_fullname_str_mv 537a2fe031a796a3bde99679ee8c24f5_***_Paul Rees
a61c15e220837ebfa52648c143769427_***_Huw Summers
author Paul Rees
Huw Summers
author2 H. D. Summers
P. Rees
J. T-W. Wang
K. T. Al-Jamal
Paul Rees
Huw Summers
format Journal article
container_title Nanoscale
container_volume 9
container_issue 20
container_start_page 6800
publishDate 2017
institution Swansea University
issn 2040-3364
2040-3372
doi_str_mv 10.1039/c7nr01561e
college_str Faculty of Science and Engineering
hierarchytype
hierarchy_top_id facultyofscienceandengineering
hierarchy_top_title Faculty of Science and Engineering
hierarchy_parent_id facultyofscienceandengineering
hierarchy_parent_title Faculty of Science and Engineering
department_str School of Engineering and Applied Sciences - Biomedical Engineering{{{_:::_}}}Faculty of Science and Engineering{{{_:::_}}}School of Engineering and Applied Sciences - Biomedical Engineering
document_store_str 1
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description The internalisation and intra-cellular distribution of carbon nanotubes (CNT) has been quantitatively assessed using imaging flow cytometry. Spatial analysis of the bright field images indicates the presence of a small sub-population (5% of cells) in which the internalised CNTs are packed into pronounced clusters, visible as dark spots due to strong optical scattering by the nanotubes. The area of these spots can be used as a label-free metric of CNT dose and we assess the relative uptake of charge-neutral CNTs, over a 24 hours exposure period across four cell types: J774 mouse macrophage cells, A549 and Calu-6 human lung cancer cells, and MCF-7 human breast cells. The relative dose as indicated by the spot-area metric closely correlates to results using the same CNT preparation, conjugated to a FITC-label and shows pronounced uptake by the J774 cells leading to a mean dose that is >60% higher than for the other cell types. Spatial evaluation of dosing clusters is also used to quantify differences in uptake by J774 cells of CNTs with different surface functionalisation. While the percentage of CNT-cluster positive cells increases from 5% to 19% when switching from charge-neutral CNTs to poly-cationic, dendron functionalised CNTs, the single cell level analysis of internalised clusters indicates a lower dose per cell of poly-cationic CNTs relative to the charge-neutral CNTs. We concluded that there is dose homeostasis i.e., the population-averaged cellular dose of CNTs remained unchanged.
published_date 2017-12-31T01:22:22Z
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