Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy

Jing Liu; Ranhua Xiong; Toon Brans; Saskia Lippens; Eef Parthoens; Francesca Cella Zanacchi; Raffaella Magrassi; Santosh K. Singh; Sreekumar Kurungot; Sabine Szunerits; Hannelore Bové; Marcel Ameloot; Juan C. Fraire; Eline Teirlinck; Sangram Keshari Samal; Riet De Rycke; Gaëlle Houthaeve; Stefaan C. De Smedt; Rabah Boukherroub; Kevin Braeckmans

doi:10.1038/s41377-018-0048-3

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Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy

Original Articles

- Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy
- Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy
- 光：科学与应用 2018年7卷第5期页码：476-485
- Affiliations：
  
  1.Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium
  2.Centre for Nano- and Biophotonics, Ghent University, Ghent, B-9000, Belgium
  3.VIB-UGent Centre for Inflammation Research, VIB, Ghent, B-9000, Belgium
  4.VIB Bioimaging Core, VIB, Ghent, B-9000, Belgium
  5.Department of Biomedical Molecular Biology, Ghent University, Ghent, B-9000, Belgium
  6.Nanophysics (NAPH), Istituto Italiano di Tecnologia, Genova, 16163, Italy
  7.Biophysics Institute (IBF), National Research Council (CNR), Via De Marini, 6-16149–GE, Genova, Italy
  8.Physical and Materials Chemistry Division, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune, 411008, India
  9.Academy of Scientific and Innovative Research, Anusandhan Bhawan, 2 RafiMarg, New Delhi, 110 001, India
  10.Univ. Lille, CNRS, Centrale Lille, ISEN, Univ. Valenciennes, UMR 8520-IEMN, Lille, F-59000, France
  11.Biomedical Research Institute, Hasselt University, Agoralaan Building C, Diepenbeek, 3590, Belgium
  12.Centre for Surface Chemistry and Catalysis, KU Leuven, Celestijnenlaan 200F, Leuven, 3001, Belgium
  13.Inflammation Research Center, Image Core Facility, VIB, 9052, Ghent, Belgium
  14.Department of Biomedical Molecular Biology, Ghent University, 9052, Ghent, Belgium
  15.Department of Biomedical Molecular Biology, Ghent University, 9052, Ghent, Belgium
  16.Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052, Gent, Belgium
  17.Univ Lille 1, Univ Lille Nord France, Lab Phys Lasers Atomes & Mol, Villeneuve Dascq, UMR 8523, 59655, France
  18.College of Chemical Engineering, Jiangsu Key Lab of Biomass-based Green Fuels and Chemicals, Nanjing Forestry University (NFU), Nanjing, 210037, China
  19.UMR 8523, Laboratoire de Physique des Lasers, Atomes et Molécules, Université de Lille, Villeneuve d'Ascq, France
  20.IEMN, UMR 8520, Université de Lille, Villeneuve d'Ascq, France
- Author bio：
  
  Kevin Braeckmans (Kevin.Braeckmans@UGent.be)
- Funds：
- DOI：10.1038/s41377-018-0048-3
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Jing Liu, Ranhua Xiong, Toon Brans, 等. Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy[J]. 光：科学与应用, 2018,7(5):476-485.

Liu, J. et al. Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy. Light: Science & Applications, 7, 476-485 (2018).
Jing Liu, Ranhua Xiong, Toon Brans, 等. Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy[J]. 光：科学与应用, 2018,7(5):476-485. DOI： 10.1038/s41377-018-0048-3.

Liu, J. et al. Repeated photoporation with graphene quantum dots enables homogeneous labeling of live cells with extrinsic markers for fluorescence microscopy. Light: Science & Applications, 7, 476-485 (2018). DOI： 10.1038/s41377-018-0048-3.

摘要

Abstract

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.

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references

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