Cargando…

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....

Descripción completa

Detalles Bibliográficos
Autores principales: Ghaemi, Reza, Tong, Justin, Gupta, Bhagwati P., Selvaganapathy, P. Ravi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
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
_version_ 1783519527146881024
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
work_keys_str_mv AT ghaemireza microfluidicdeviceformicroinjectionofcaenorhabditiselegans
AT tongjustin microfluidicdeviceformicroinjectionofcaenorhabditiselegans
AT guptabhagwatip microfluidicdeviceformicroinjectionofcaenorhabditiselegans
AT selvaganapathypravi microfluidicdeviceformicroinjectionofcaenorhabditiselegans