Abstract
The cell membrane is crucial for cell survival, and ensuring its integrity is essential
as the cell experiences injuries throughout its entire life cycle. To prevent damage to
the membrane, cells have developed efficient plasma membrane repair mechanisms.
These repair mechanisms can be studied by combining confocal microscopy and
nanoscale thermoplasmonics to identify and investigate the role of key proteins, such
as annexins, involved in surface repair in living cells and membrane model systems.
The puncturing method employs a laser to induce highly localized heating upon
nanoparticle irradiation. The use of near-infrared light minimizes phototoxicity in the
biological sample, while the majority of the absorption takes place in the near-infrared
resonant plasmonic nanoparticle. This thermoplasmonic method has been exploited
for potential photothermal and biophysical research to enhance the understanding
of intracellular mechanisms and cellular responses through vesicle and cell fusion
studies. The approach has shown to be complementary to existing methods for
membrane disruption, such as mechanically, chemically, or optically induced injuries,
and provides a high level of control by inflicting extremely localized injuries. The extent
of the injury is limited to the vicinity of the spherical nanoparticle, and no detrimental
damage occurs along the beam path as opposed to pulsed lasers using different
wavelengths. Despite certain limitations, such as the formation of nanobubbles, the
thermoplasmonic method offers a unique tool for investigating cellular responses in
plasma membrane repair in an almost native environment without compromising cell
viability.
When integrated with confocal microscopy, the puncturing method can provide a
mechanistic understanding of membrane dynamics in model membrane systems as
well as quantitative information on protein responses to membrane damage, including
protein recruitment and their biophysical function. Overall, the application of this
Copyright © 2024 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
License
jove.com January 2024 • 203 • e65776 • Page 2 of 21
method to reduced model systems can enhance our understanding of the intricate
plasma membrane repair machinery in living cells.
as the cell experiences injuries throughout its entire life cycle. To prevent damage to
the membrane, cells have developed efficient plasma membrane repair mechanisms.
These repair mechanisms can be studied by combining confocal microscopy and
nanoscale thermoplasmonics to identify and investigate the role of key proteins, such
as annexins, involved in surface repair in living cells and membrane model systems.
The puncturing method employs a laser to induce highly localized heating upon
nanoparticle irradiation. The use of near-infrared light minimizes phototoxicity in the
biological sample, while the majority of the absorption takes place in the near-infrared
resonant plasmonic nanoparticle. This thermoplasmonic method has been exploited
for potential photothermal and biophysical research to enhance the understanding
of intracellular mechanisms and cellular responses through vesicle and cell fusion
studies. The approach has shown to be complementary to existing methods for
membrane disruption, such as mechanically, chemically, or optically induced injuries,
and provides a high level of control by inflicting extremely localized injuries. The extent
of the injury is limited to the vicinity of the spherical nanoparticle, and no detrimental
damage occurs along the beam path as opposed to pulsed lasers using different
wavelengths. Despite certain limitations, such as the formation of nanobubbles, the
thermoplasmonic method offers a unique tool for investigating cellular responses in
plasma membrane repair in an almost native environment without compromising cell
viability.
When integrated with confocal microscopy, the puncturing method can provide a
mechanistic understanding of membrane dynamics in model membrane systems as
well as quantitative information on protein responses to membrane damage, including
protein recruitment and their biophysical function. Overall, the application of this
Copyright © 2024 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
License
jove.com January 2024 • 203 • e65776 • Page 2 of 21
method to reduced model systems can enhance our understanding of the intricate
plasma membrane repair machinery in living cells.
| Original language | English |
|---|---|
| Article number | e65776 |
| Number of pages | 21 |
| Journal | JoVe journal |
| Volume | 203 |
| Publication status | Published - 2024 |
| Externally published | Yes |