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Microfluidic Device for Microinjection of Caenorhabditis elegans
Microinjection is an established and reliable method to deliver transgenic constructs and other reagents to specific locations in C. elegans worms. Specifically, microinjection of a desired DNA construct into the distal gonad is the most widely used method to generate germ-line transformation of C....
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7143065/ https://www.ncbi.nlm.nih.gov/pubmed/32168862 http://dx.doi.org/10.3390/mi11030295 |
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author | Ghaemi, Reza Tong, Justin Gupta, Bhagwati P. Selvaganapathy, P. Ravi |
author_facet | Ghaemi, Reza Tong, Justin Gupta, Bhagwati P. Selvaganapathy, P. Ravi |
author_sort | Ghaemi, Reza |
collection | PubMed |
description | Microinjection is an established and reliable method to deliver transgenic constructs and other reagents to specific locations in C. elegans worms. Specifically, microinjection of a desired DNA construct into the distal gonad is the most widely used method to generate germ-line transformation of C. elegans. Although, current C. elegans microinjection method is effective to produce transgenic worms, it requires expensive multi degree of freedom (DOF) micromanipulator, careful injection alignment procedure and skilled operator, all of which make it slow and not suitable for scaling to high throughput. A few microfabricated microinjectors have been developed recently to address these issues. However, none of them are capable of immobilizing a freely mobile animal such as C. elegans worm using a passive immobilization mechanism. Here, a microfluidic microinjector was developed to passively immobilize a freely mobile animal such as C. elegans and simultaneously perform microinjection by using a simple and fast mechanism for needle actuation. The entire process of the microinjection takes ~30 s which includes 10 s for worm loading and aligning, 5 s needle penetration, 5 s reagent injection and 5 s worm unloading. The device is suitable for high-throughput and can be potentially used for creating transgenic C. elegans. |
format | Online Article Text |
id | pubmed-7143065 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-71430652020-04-14 Microfluidic Device for Microinjection of Caenorhabditis elegans Ghaemi, Reza Tong, Justin Gupta, Bhagwati P. Selvaganapathy, P. Ravi Micromachines (Basel) Article Microinjection is an established and reliable method to deliver transgenic constructs and other reagents to specific locations in C. elegans worms. Specifically, microinjection of a desired DNA construct into the distal gonad is the most widely used method to generate germ-line transformation of C. elegans. Although, current C. elegans microinjection method is effective to produce transgenic worms, it requires expensive multi degree of freedom (DOF) micromanipulator, careful injection alignment procedure and skilled operator, all of which make it slow and not suitable for scaling to high throughput. A few microfabricated microinjectors have been developed recently to address these issues. However, none of them are capable of immobilizing a freely mobile animal such as C. elegans worm using a passive immobilization mechanism. Here, a microfluidic microinjector was developed to passively immobilize a freely mobile animal such as C. elegans and simultaneously perform microinjection by using a simple and fast mechanism for needle actuation. The entire process of the microinjection takes ~30 s which includes 10 s for worm loading and aligning, 5 s needle penetration, 5 s reagent injection and 5 s worm unloading. The device is suitable for high-throughput and can be potentially used for creating transgenic C. elegans. MDPI 2020-03-11 /pmc/articles/PMC7143065/ /pubmed/32168862 http://dx.doi.org/10.3390/mi11030295 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ghaemi, Reza Tong, Justin Gupta, Bhagwati P. Selvaganapathy, P. Ravi Microfluidic Device for Microinjection of Caenorhabditis elegans |
title | Microfluidic Device for Microinjection of Caenorhabditis elegans |
title_full | Microfluidic Device for Microinjection of Caenorhabditis elegans |
title_fullStr | Microfluidic Device for Microinjection of Caenorhabditis elegans |
title_full_unstemmed | Microfluidic Device for Microinjection of Caenorhabditis elegans |
title_short | Microfluidic Device for Microinjection of Caenorhabditis elegans |
title_sort | microfluidic device for microinjection of caenorhabditis elegans |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7143065/ https://www.ncbi.nlm.nih.gov/pubmed/32168862 http://dx.doi.org/10.3390/mi11030295 |
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