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A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans

Femtosecond laser nanosurgery has been widely accepted as an axonal injury model, enabling nerve regeneration studies in the small model organism, Caenorhabditis elegans. To overcome the time limitations of manual worm handling techniques, automation and new immobilization technologies must be adopt...

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Autores principales: Gokce, Sertan Kutal, Guo, Samuel X., Ghorashian, Navid, Everett, W. Neil, Jarrell, Travis, Kottek, Aubri, Bovik, Alan C., Ben-Yakar, Adela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4254741/
https://www.ncbi.nlm.nih.gov/pubmed/25470130
http://dx.doi.org/10.1371/journal.pone.0113917
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author Gokce, Sertan Kutal
Guo, Samuel X.
Ghorashian, Navid
Everett, W. Neil
Jarrell, Travis
Kottek, Aubri
Bovik, Alan C.
Ben-Yakar, Adela
author_facet Gokce, Sertan Kutal
Guo, Samuel X.
Ghorashian, Navid
Everett, W. Neil
Jarrell, Travis
Kottek, Aubri
Bovik, Alan C.
Ben-Yakar, Adela
author_sort Gokce, Sertan Kutal
collection PubMed
description Femtosecond laser nanosurgery has been widely accepted as an axonal injury model, enabling nerve regeneration studies in the small model organism, Caenorhabditis elegans. To overcome the time limitations of manual worm handling techniques, automation and new immobilization technologies must be adopted to improve throughput in these studies. While new microfluidic immobilization techniques have been developed that promise to reduce the time required for axotomies, there is a need for automated procedures to minimize the required amount of human intervention and accelerate the axotomy processes crucial for high-throughput. Here, we report a fully automated microfluidic platform for performing laser axotomies of fluorescently tagged neurons in living Caenorhabditis elegans. The presented automation process reduces the time required to perform axotomies within individual worms to ∼17 s/worm, at least one order of magnitude faster than manual approaches. The full automation is achieved with a unique chip design and an operation sequence that is fully computer controlled and synchronized with efficient and accurate image processing algorithms. The microfluidic device includes a T-shaped architecture and three-dimensional microfluidic interconnects to serially transport, position, and immobilize worms. The image processing algorithms can identify and precisely position axons targeted for ablation. There were no statistically significant differences observed in reconnection probabilities between axotomies carried out with the automated system and those performed manually with anesthetics. The overall success rate of automated axotomies was 67.4±3.2% of the cases (236/350) at an average processing rate of 17.0±2.4 s. This fully automated platform establishes a promising methodology for prospective genome-wide screening of nerve regeneration in C. elegans in a truly high-throughput manner.
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spelling pubmed-42547412014-12-11 A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans Gokce, Sertan Kutal Guo, Samuel X. Ghorashian, Navid Everett, W. Neil Jarrell, Travis Kottek, Aubri Bovik, Alan C. Ben-Yakar, Adela PLoS One Research Article Femtosecond laser nanosurgery has been widely accepted as an axonal injury model, enabling nerve regeneration studies in the small model organism, Caenorhabditis elegans. To overcome the time limitations of manual worm handling techniques, automation and new immobilization technologies must be adopted to improve throughput in these studies. While new microfluidic immobilization techniques have been developed that promise to reduce the time required for axotomies, there is a need for automated procedures to minimize the required amount of human intervention and accelerate the axotomy processes crucial for high-throughput. Here, we report a fully automated microfluidic platform for performing laser axotomies of fluorescently tagged neurons in living Caenorhabditis elegans. The presented automation process reduces the time required to perform axotomies within individual worms to ∼17 s/worm, at least one order of magnitude faster than manual approaches. The full automation is achieved with a unique chip design and an operation sequence that is fully computer controlled and synchronized with efficient and accurate image processing algorithms. The microfluidic device includes a T-shaped architecture and three-dimensional microfluidic interconnects to serially transport, position, and immobilize worms. The image processing algorithms can identify and precisely position axons targeted for ablation. There were no statistically significant differences observed in reconnection probabilities between axotomies carried out with the automated system and those performed manually with anesthetics. The overall success rate of automated axotomies was 67.4±3.2% of the cases (236/350) at an average processing rate of 17.0±2.4 s. This fully automated platform establishes a promising methodology for prospective genome-wide screening of nerve regeneration in C. elegans in a truly high-throughput manner. Public Library of Science 2014-12-03 /pmc/articles/PMC4254741/ /pubmed/25470130 http://dx.doi.org/10.1371/journal.pone.0113917 Text en © 2014 Gokce et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Gokce, Sertan Kutal
Guo, Samuel X.
Ghorashian, Navid
Everett, W. Neil
Jarrell, Travis
Kottek, Aubri
Bovik, Alan C.
Ben-Yakar, Adela
A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title_full A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title_fullStr A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title_full_unstemmed A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title_short A Fully Automated Microfluidic Femtosecond Laser Axotomy Platform for Nerve Regeneration Studies in C. elegans
title_sort fully automated microfluidic femtosecond laser axotomy platform for nerve regeneration studies in c. elegans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4254741/
https://www.ncbi.nlm.nih.gov/pubmed/25470130
http://dx.doi.org/10.1371/journal.pone.0113917
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