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Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy
In the replacement of genetic probes, there is increasing interest in labeling living cells with high-quality extrinsic labels, which avoid over-expression artifacts and are available in a wide spectral range. This calls for a broadly applicable technology that can deliver such labels unambiguously...
Autores principales: | , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6106998/ https://www.ncbi.nlm.nih.gov/pubmed/30839577 http://dx.doi.org/10.1038/s41377-018-0048-3 |
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author | Liu, Jing Xiong, Ranhua Brans, Toon Lippens, Saskia Parthoens, Eef Zanacchi, Francesca Cella Magrassi, Raffaella Singh, Santosh K. Kurungot, Sreekumar Szunerits, Sabine Bové, Hannelore Ameloot, Marcel Fraire, Juan C. Teirlinck, Eline Samal, Sangram Keshari Rycke, Riet De Houthaeve, Gaëlle De Smedt, Stefaan C. Boukherroub, Rabah Braeckmans, Kevin |
author_facet | Liu, Jing Xiong, Ranhua Brans, Toon Lippens, Saskia Parthoens, Eef Zanacchi, Francesca Cella Magrassi, Raffaella Singh, Santosh K. Kurungot, Sreekumar Szunerits, Sabine Bové, Hannelore Ameloot, Marcel Fraire, Juan C. Teirlinck, Eline Samal, Sangram Keshari Rycke, Riet De Houthaeve, Gaëlle De Smedt, Stefaan C. Boukherroub, Rabah Braeckmans, Kevin |
author_sort | Liu, Jing |
collection | PubMed |
description | In the replacement of genetic probes, there is increasing interest in labeling living cells with high-quality extrinsic labels, which avoid over-expression artifacts and are available in a wide spectral range. This calls for a broadly applicable technology that can deliver such labels unambiguously to the cytosol of living cells. Here, we demonstrate that nanoparticle-sensitized photoporation can be used to this end as an emerging intracellular delivery technique. We replace the traditionally used gold nanoparticles with graphene nanoparticles as photothermal sensitizers to permeabilize the cell membrane upon laser irradiation. We demonstrate that the enhanced thermal stability of graphene quantum dots allows the formation of multiple vapor nanobubbles upon irradiation with short laser pulses, allowing the delivery of a variety of extrinsic cell labels efficiently and homogeneously into live cells. We demonstrate high-quality time-lapse imaging with confocal, total internal reflection fluorescence (TIRF), and Airyscan super-resolution microscopy. As the entire procedure is readily compatible with fluorescence (super resolution) microscopy, photoporation with graphene quantum dots has the potential to become the long-awaited generic platform for controlled intracellular delivery of fluorescent labels for live-cell imaging. |
format | Online Article Text |
id | pubmed-6106998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-61069982018-08-30 Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy Liu, Jing Xiong, Ranhua Brans, Toon Lippens, Saskia Parthoens, Eef Zanacchi, Francesca Cella Magrassi, Raffaella Singh, Santosh K. Kurungot, Sreekumar Szunerits, Sabine Bové, Hannelore Ameloot, Marcel Fraire, Juan C. Teirlinck, Eline Samal, Sangram Keshari Rycke, Riet De Houthaeve, Gaëlle De Smedt, Stefaan C. Boukherroub, Rabah Braeckmans, Kevin Light Sci Appl Article In the replacement of genetic probes, there is increasing interest in labeling living cells with high-quality extrinsic labels, which avoid over-expression artifacts and are available in a wide spectral range. This calls for a broadly applicable technology that can deliver such labels unambiguously to the cytosol of living cells. Here, we demonstrate that nanoparticle-sensitized photoporation can be used to this end as an emerging intracellular delivery technique. We replace the traditionally used gold nanoparticles with graphene nanoparticles as photothermal sensitizers to permeabilize the cell membrane upon laser irradiation. We demonstrate that the enhanced thermal stability of graphene quantum dots allows the formation of multiple vapor nanobubbles upon irradiation with short laser pulses, allowing the delivery of a variety of extrinsic cell labels efficiently and homogeneously into live cells. We demonstrate high-quality time-lapse imaging with confocal, total internal reflection fluorescence (TIRF), and Airyscan super-resolution microscopy. As the entire procedure is readily compatible with fluorescence (super resolution) microscopy, photoporation with graphene quantum dots has the potential to become the long-awaited generic platform for controlled intracellular delivery of fluorescent labels for live-cell imaging. Nature Publishing Group UK 2018-08-08 /pmc/articles/PMC6106998/ /pubmed/30839577 http://dx.doi.org/10.1038/s41377-018-0048-3 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Liu, Jing Xiong, Ranhua Brans, Toon Lippens, Saskia Parthoens, Eef Zanacchi, Francesca Cella Magrassi, Raffaella Singh, Santosh K. Kurungot, Sreekumar Szunerits, Sabine Bové, Hannelore Ameloot, Marcel Fraire, Juan C. Teirlinck, Eline Samal, Sangram Keshari Rycke, Riet De Houthaeve, Gaëlle De Smedt, Stefaan C. Boukherroub, Rabah Braeckmans, Kevin Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title | Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title_full | Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title_fullStr | Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title_full_unstemmed | Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title_short | Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
title_sort | repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6106998/ https://www.ncbi.nlm.nih.gov/pubmed/30839577 http://dx.doi.org/10.1038/s41377-018-0048-3 |
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