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Nanostraw-Assisted Cellular Injection of Fluorescent Nanodiamonds via Direct Membrane Opening

Hebisch, Elke LU ; Hjort, Martin LU orcid ; Volpati, Diogo LU and Prinz, Christelle N. LU (2021) In Small 17(7).
Abstract

Due to their stable fluorescence, biocompatibility, and amenability to functionalization, fluorescent nanodiamonds (FND) are promising materials for long term cell labeling and tracking. However, transporting them to the cytosol remains a major challenge, due to low internalization efficiencies and endosomal entrapment. Here, nanostraws in combination with low voltage electroporation pulses are used to achieve direct delivery of FND to the cytosol. The nanostraw delivery leads to efficient and rapid FND transport into cells compared to when incubating cells in a FND-containing medium. Moreover, whereas all internalized FND delivered by incubation end up in lysosomes, a significantly larger proportion of nanostraw-injected FND are in the... (More)

Due to their stable fluorescence, biocompatibility, and amenability to functionalization, fluorescent nanodiamonds (FND) are promising materials for long term cell labeling and tracking. However, transporting them to the cytosol remains a major challenge, due to low internalization efficiencies and endosomal entrapment. Here, nanostraws in combination with low voltage electroporation pulses are used to achieve direct delivery of FND to the cytosol. The nanostraw delivery leads to efficient and rapid FND transport into cells compared to when incubating cells in a FND-containing medium. Moreover, whereas all internalized FND delivered by incubation end up in lysosomes, a significantly larger proportion of nanostraw-injected FND are in the cytosol, which opens up for using FND as cellular probes. Furthermore, in order to answer the long-standing question in the field of nano-biology regarding the state of the cell membrane on hollow nanostructures, live cell stimulated emission depletion (STED) microscopy is performed to image directly the state of the membrane on nanostraws. The time-lapse STED images reveal that the cell membrane opens entirely on top of nanostraws upon application of gentle electrical pulses, which supports the hypothesis that many FND are delivered directly to the cytosol, avoiding endocytosis and lysosomal entrapment.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cell transfection, electroporation, nanodiamonds, nanostraws, STED microscopy
in
Small
volume
17
issue
7
article number
2006421
publisher
John Wiley & Sons Inc.
external identifiers
  • scopus:85099738683
  • pmid:33502091
ISSN
1613-6810
DOI
10.1002/smll.202006421
language
English
LU publication?
yes
id
e8d0bba8-8bf5-485c-ba51-52f0e510225d
date added to LUP
2021-02-02 14:34:07
date last changed
2024-06-27 07:56:53
@article{e8d0bba8-8bf5-485c-ba51-52f0e510225d,
  abstract     = {{<p>Due to their stable fluorescence, biocompatibility, and amenability to functionalization, fluorescent nanodiamonds (FND) are promising materials for long term cell labeling and tracking. However, transporting them to the cytosol remains a major challenge, due to low internalization efficiencies and endosomal entrapment. Here, nanostraws in combination with low voltage electroporation pulses are used to achieve direct delivery of FND to the cytosol. The nanostraw delivery leads to efficient and rapid FND transport into cells compared to when incubating cells in a FND-containing medium. Moreover, whereas all internalized FND delivered by incubation end up in lysosomes, a significantly larger proportion of nanostraw-injected FND are in the cytosol, which opens up for using FND as cellular probes. Furthermore, in order to answer the long-standing question in the field of nano-biology regarding the state of the cell membrane on hollow nanostructures, live cell stimulated emission depletion (STED) microscopy is performed to image directly the state of the membrane on nanostraws. The time-lapse STED images reveal that the cell membrane opens entirely on top of nanostraws upon application of gentle electrical pulses, which supports the hypothesis that many FND are delivered directly to the cytosol, avoiding endocytosis and lysosomal entrapment.</p>}},
  author       = {{Hebisch, Elke and Hjort, Martin and Volpati, Diogo and Prinz, Christelle N.}},
  issn         = {{1613-6810}},
  keywords     = {{cell transfection; electroporation; nanodiamonds; nanostraws; STED microscopy}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{7}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Small}},
  title        = {{Nanostraw-Assisted Cellular Injection of Fluorescent Nanodiamonds via Direct Membrane Opening}},
  url          = {{http://dx.doi.org/10.1002/smll.202006421}},
  doi          = {{10.1002/smll.202006421}},
  volume       = {{17}},
  year         = {{2021}},
}