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Spinal cord injury in zebrafish induced by near-infrared femtosecond laser pulses

Dehnisch Ellström, Ivar ; Spulber, Stefan ; Hultin, Sara ; Norlin, Nils LU ; Ceccatelli, Sandra ; Hultling, Claes and Uhlén, Per (2019) In Journal of Neuroscience Methods 311. p.259-266
Abstract

BACKGROUND: The spinal cord is composed of a large number of cells that interact to allow the organism to function. To perform detail studies of cellular processes involved in spinal cord injury (SCI), one must use repeatable and specific methods to target and injure restricted areas of the spinal cord.

NEW METHOD: We propose a robust method to induce SCI in zebrafish by laser light. With a 2-photon microscope equipped with a femtosecond near-infrared pump laser, we explored the effects of laser beam exposure time, area, and intensity to induce precise and repeatable SCI with minimized collateral damage to neighboring cells.

RESULTS: Through behavioral studies in zebrafish larvae, we assessed the functional outcome of... (More)

BACKGROUND: The spinal cord is composed of a large number of cells that interact to allow the organism to function. To perform detail studies of cellular processes involved in spinal cord injury (SCI), one must use repeatable and specific methods to target and injure restricted areas of the spinal cord.

NEW METHOD: We propose a robust method to induce SCI in zebrafish by laser light. With a 2-photon microscope equipped with a femtosecond near-infrared pump laser, we explored the effects of laser beam exposure time, area, and intensity to induce precise and repeatable SCI with minimized collateral damage to neighboring cells.

RESULTS: Through behavioral studies in zebrafish larvae, we assessed the functional outcome of intensive laser light directed at the spinal cord. Our experiments revealed that a laser pulse with wavelength 800 nm, duration 2.6 ms, and light intensity 390 mW was sufficient to induce controlled cell death in a single cell or a spinal cord segment. Collateral damage was observed if cells were exposed to laser pulses exceeding 470 mW. With these settings, we could induce precise and repeatable SCI in zebrafish larvae, resulting in loss of motor and sensory function.

COMPARISON WITH EXISTING METHOD(S): Our method offers a simple and more controlled setting to induce SCI in zebrafish. We describe how the near-infrared femtosecond laser should be adjusted for achieving optimal results with minimal collateral damage.

CONCLUSIONS: We present a precise and robust method for inducing SCI in zebrafish with single-cell resolution using femtosecond near-infrared laser pulses.

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author
; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Animals, Disease Models, Animal, Larva, Lasers, Locomotion, Microscopy, Confocal/methods, Microsurgery/instrumentation, Neurosurgical Procedures/instrumentation, Spinal Cord Injuries/pathology, Zebrafish
in
Journal of Neuroscience Methods
volume
311
pages
8 pages
publisher
Elsevier
external identifiers
  • pmid:30389486
  • scopus:85056003198
ISSN
1872-678X
DOI
10.1016/j.jneumeth.2018.10.035
language
English
LU publication?
no
additional info
Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.
id
20c7180b-6fcd-423d-b632-64e1f81d94ef
date added to LUP
2020-09-09 18:32:28
date last changed
2024-03-05 10:31:27
@article{20c7180b-6fcd-423d-b632-64e1f81d94ef,
  abstract     = {{<p>BACKGROUND: The spinal cord is composed of a large number of cells that interact to allow the organism to function. To perform detail studies of cellular processes involved in spinal cord injury (SCI), one must use repeatable and specific methods to target and injure restricted areas of the spinal cord.</p><p>NEW METHOD: We propose a robust method to induce SCI in zebrafish by laser light. With a 2-photon microscope equipped with a femtosecond near-infrared pump laser, we explored the effects of laser beam exposure time, area, and intensity to induce precise and repeatable SCI with minimized collateral damage to neighboring cells.</p><p>RESULTS: Through behavioral studies in zebrafish larvae, we assessed the functional outcome of intensive laser light directed at the spinal cord. Our experiments revealed that a laser pulse with wavelength 800 nm, duration 2.6 ms, and light intensity 390 mW was sufficient to induce controlled cell death in a single cell or a spinal cord segment. Collateral damage was observed if cells were exposed to laser pulses exceeding 470 mW. With these settings, we could induce precise and repeatable SCI in zebrafish larvae, resulting in loss of motor and sensory function.</p><p>COMPARISON WITH EXISTING METHOD(S): Our method offers a simple and more controlled setting to induce SCI in zebrafish. We describe how the near-infrared femtosecond laser should be adjusted for achieving optimal results with minimal collateral damage.</p><p>CONCLUSIONS: We present a precise and robust method for inducing SCI in zebrafish with single-cell resolution using femtosecond near-infrared laser pulses.</p>}},
  author       = {{Dehnisch Ellström, Ivar and Spulber, Stefan and Hultin, Sara and Norlin, Nils and Ceccatelli, Sandra and Hultling, Claes and Uhlén, Per}},
  issn         = {{1872-678X}},
  keywords     = {{Animals; Disease Models, Animal; Larva; Lasers; Locomotion; Microscopy, Confocal/methods; Microsurgery/instrumentation; Neurosurgical Procedures/instrumentation; Spinal Cord Injuries/pathology; Zebrafish}},
  language     = {{eng}},
  month        = {{01}},
  pages        = {{259--266}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Neuroscience Methods}},
  title        = {{Spinal cord injury in zebrafish induced by near-infrared femtosecond laser pulses}},
  url          = {{http://dx.doi.org/10.1016/j.jneumeth.2018.10.035}},
  doi          = {{10.1016/j.jneumeth.2018.10.035}},
  volume       = {{311}},
  year         = {{2019}},
}