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Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays

New markers are constantly emerging that identify smaller and smaller subpopulations of immune cells. However, there is a growing awareness that even within very small populations, there is a marked functional heterogeneity and that measurements at the population level only gives an average estimate...

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Autores principales: Guldevall, Karolin, Vanherberghen, Bruno, Frisk, Thomas, Hurtig, Johan, Christakou, Athanasia E., Manneberg, Otto, Lindström, Sara, Andersson-Svahn, Helene, Wiklund, Martin, Önfelt, Björn
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2980494/
https://www.ncbi.nlm.nih.gov/pubmed/21103395
http://dx.doi.org/10.1371/journal.pone.0015453
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author Guldevall, Karolin
Vanherberghen, Bruno
Frisk, Thomas
Hurtig, Johan
Christakou, Athanasia E.
Manneberg, Otto
Lindström, Sara
Andersson-Svahn, Helene
Wiklund, Martin
Önfelt, Björn
author_facet Guldevall, Karolin
Vanherberghen, Bruno
Frisk, Thomas
Hurtig, Johan
Christakou, Athanasia E.
Manneberg, Otto
Lindström, Sara
Andersson-Svahn, Helene
Wiklund, Martin
Önfelt, Björn
author_sort Guldevall, Karolin
collection PubMed
description New markers are constantly emerging that identify smaller and smaller subpopulations of immune cells. However, there is a growing awareness that even within very small populations, there is a marked functional heterogeneity and that measurements at the population level only gives an average estimate of the behaviour of that pool of cells. New techniques to analyze single immune cells over time are needed to overcome this limitation. For that purpose, we have designed and evaluated microwell array systems made from two materials, polydimethylsiloxane (PDMS) and silicon, for high-resolution imaging of individual natural killer (NK) cell responses. Both materials were suitable for short-term studies (<4 hours) but only silicon wells allowed long-term studies (several days). Time-lapse imaging of NK cell cytotoxicity in these microwell arrays revealed that roughly 30% of the target cells died much more rapidly than the rest upon NK cell encounter. This unexpected heterogeneity may reflect either separate mechanisms of killing or different killing efficiency by individual NK cells. Furthermore, we show that high-resolution imaging of inhibitory synapse formation, defined by clustering of MHC class I at the interface between NK and target cells, is possible in these microwells. We conclude that live cell imaging of NK-target cell interactions in multi-well microstructures are possible. The technique enables novel types of assays and allow data collection at a level of resolution not previously obtained. Furthermore, due to the large number of wells that can be simultaneously imaged, new statistical information is obtained that will lead to a better understanding of the function and regulation of the immune system at the single cell level.
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spelling pubmed-29804942010-11-22 Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays Guldevall, Karolin Vanherberghen, Bruno Frisk, Thomas Hurtig, Johan Christakou, Athanasia E. Manneberg, Otto Lindström, Sara Andersson-Svahn, Helene Wiklund, Martin Önfelt, Björn PLoS One Research Article New markers are constantly emerging that identify smaller and smaller subpopulations of immune cells. However, there is a growing awareness that even within very small populations, there is a marked functional heterogeneity and that measurements at the population level only gives an average estimate of the behaviour of that pool of cells. New techniques to analyze single immune cells over time are needed to overcome this limitation. For that purpose, we have designed and evaluated microwell array systems made from two materials, polydimethylsiloxane (PDMS) and silicon, for high-resolution imaging of individual natural killer (NK) cell responses. Both materials were suitable for short-term studies (<4 hours) but only silicon wells allowed long-term studies (several days). Time-lapse imaging of NK cell cytotoxicity in these microwell arrays revealed that roughly 30% of the target cells died much more rapidly than the rest upon NK cell encounter. This unexpected heterogeneity may reflect either separate mechanisms of killing or different killing efficiency by individual NK cells. Furthermore, we show that high-resolution imaging of inhibitory synapse formation, defined by clustering of MHC class I at the interface between NK and target cells, is possible in these microwells. We conclude that live cell imaging of NK-target cell interactions in multi-well microstructures are possible. The technique enables novel types of assays and allow data collection at a level of resolution not previously obtained. Furthermore, due to the large number of wells that can be simultaneously imaged, new statistical information is obtained that will lead to a better understanding of the function and regulation of the immune system at the single cell level. Public Library of Science 2010-11-12 /pmc/articles/PMC2980494/ /pubmed/21103395 http://dx.doi.org/10.1371/journal.pone.0015453 Text en Guldevall et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Guldevall, Karolin
Vanherberghen, Bruno
Frisk, Thomas
Hurtig, Johan
Christakou, Athanasia E.
Manneberg, Otto
Lindström, Sara
Andersson-Svahn, Helene
Wiklund, Martin
Önfelt, Björn
Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title_full Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title_fullStr Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title_full_unstemmed Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title_short Imaging Immune Surveillance of Individual Natural Killer Cells Confined in Microwell Arrays
title_sort imaging immune surveillance of individual natural killer cells confined in microwell arrays
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2980494/
https://www.ncbi.nlm.nih.gov/pubmed/21103395
http://dx.doi.org/10.1371/journal.pone.0015453
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