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...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 superresolution 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.展开更多
基金support from the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program(grant agreement No 648124)from the Ghent University Special Research Fund(01B04912)with gratitude+8 种基金support from the China Scholarship Council(CSC)(201506750012)the Special Research Fund from Ghent University(01SC1416)support from the China Scholarship Council(CSC)(2010634103)from the Research Foundation Flanders(Fonds Wetenschappelijk Onderzoek,FWO)for a doctoral fellowship(11ZB115N)from the Agency for Innovation by Science and Technology(IWT)support from the Centre National de la Recherche Scientifique(CNRS),the University of Lille,the Hauts-de-France region,the CPER“Photonics for Society”the EU union through FLAG-ERA JTC 2015-Graphtivitythe Marie Sklodowska-Curie action(H2020-MSCA-RISE-2015,PANG-690836)support by the FWO Research Community“Scanning and Wide Field Microscopy of(Bio)-organic Systems”and the Province of Limburg(Belgium)for the financial support within the tUL IMPULS FASE II program。
文摘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 superresolution 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.
