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Live single-cell laser tag
The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria su...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4876456/ https://www.ncbi.nlm.nih.gov/pubmed/27198043 http://dx.doi.org/10.1038/ncomms11636 |
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author | Binan, Loïc Mazzaferri, Javier Choquet, Karine Lorenzo, Louis-Etienne Wang, Yu Chang Affar, El Bachir De Koninck, Yves Ragoussis, Jiannis Kleinman, Claudia L. Costantino, Santiago |
author_facet | Binan, Loïc Mazzaferri, Javier Choquet, Karine Lorenzo, Louis-Etienne Wang, Yu Chang Affar, El Bachir De Koninck, Yves Ragoussis, Jiannis Kleinman, Claudia L. Costantino, Santiago |
author_sort | Binan, Loïc |
collection | PubMed |
description | The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types. |
format | Online Article Text |
id | pubmed-4876456 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-48764562016-06-02 Live single-cell laser tag Binan, Loïc Mazzaferri, Javier Choquet, Karine Lorenzo, Louis-Etienne Wang, Yu Chang Affar, El Bachir De Koninck, Yves Ragoussis, Jiannis Kleinman, Claudia L. Costantino, Santiago Nat Commun Article The ability to conduct image-based, non-invasive cell tagging, independent of genetic engineering, is key to cell biology applications. Here we introduce cell labelling via photobleaching (CLaP), a method that enables instant, specific tagging of individual cells based on a wide array of criteria such as shape, behaviour or positional information. CLaP uses laser illumination to crosslink biotin onto the plasma membrane, coupled with streptavidin conjugates to label individual cells for genomic, cell-tracking, flow cytometry or ultra-microscopy applications. We show that the incorporated mark is stable, non-toxic, retained for several days, and transferred by cell division but not to adjacent cells in culture. To demonstrate the potential of CLaP for genomic applications, we combine CLaP with microfluidics-based single-cell capture followed by transcriptome-wide next-generation sequencing. Finally, we show that CLaP can also be exploited for inducing transient cell adhesion to substrates for microengineering cultures with spatially patterned cell types. Nature Publishing Group 2016-05-20 /pmc/articles/PMC4876456/ /pubmed/27198043 http://dx.doi.org/10.1038/ncomms11636 Text en Copyright © 2016, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Binan, Loïc Mazzaferri, Javier Choquet, Karine Lorenzo, Louis-Etienne Wang, Yu Chang Affar, El Bachir De Koninck, Yves Ragoussis, Jiannis Kleinman, Claudia L. Costantino, Santiago Live single-cell laser tag |
title | Live single-cell laser tag |
title_full | Live single-cell laser tag |
title_fullStr | Live single-cell laser tag |
title_full_unstemmed | Live single-cell laser tag |
title_short | Live single-cell laser tag |
title_sort | live single-cell laser tag |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4876456/ https://www.ncbi.nlm.nih.gov/pubmed/27198043 http://dx.doi.org/10.1038/ncomms11636 |
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