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Preparation of neuronal co-cultures with single cell precision

Preparation of neuronal co-cultures with single cell precision
Preparation of neuronal co-cultures with single cell precision
Microfluidic embodiments of the Campenot chamber have attracted great interest from the neuroscience community. These interconnected co-culture platforms can be used to
investigate a variety of questions, spanning developmental and functional neurobiology to infection and disease propagation. However, conventional systems require significant cellular inputs (many thousands per compartment), inadequate for studying low abundance cells, such as primary dopaminergic substantia nigra, spiral ganglia and Drosophilia melanogaster neurons, and impractical for high throughput experimentation. The dense cultures are also highly locally entangled, with few outgrowths (<10%) interconnecting the two cultures. In this paper straightforward microfluidic and patterning protocols are described which address these challenges: (i) a microfluidic single neuron arraying method, and (ii) a water masking method for plasma patterning biomaterial coatings to register neurons and promote outgrowth between compartments. Minimalistic neuronal co-cultures were prepared with high-level (>85%) inter-compartment connectivity and can be used for high throughput neurobiology experiments with single cell precision
1940-087X
e51389
Dinh, Ngoc-Duy
caa5d6c7-79aa-47c4-9612-a54898102fb8
Chiang, Ya-Yu
0ab4fe9e-9a3e-4594-a13c-73f68c13b8b3
Hardelauf, Heike
ff18e0f8-62ed-4edf-9746-f72701657440
Waide, Sarah
ccca6a91-1339-435e-ac06-5ef9127cb288
Janasek, Dirk
39a248d4-ac4a-4f07-9919-7bc1098cd715
West, Jonathan
f1c2e060-16c3-44c0-af70-242a1c58b968
Dinh, Ngoc-Duy
caa5d6c7-79aa-47c4-9612-a54898102fb8
Chiang, Ya-Yu
0ab4fe9e-9a3e-4594-a13c-73f68c13b8b3
Hardelauf, Heike
ff18e0f8-62ed-4edf-9746-f72701657440
Waide, Sarah
ccca6a91-1339-435e-ac06-5ef9127cb288
Janasek, Dirk
39a248d4-ac4a-4f07-9919-7bc1098cd715
West, Jonathan
f1c2e060-16c3-44c0-af70-242a1c58b968

Dinh, Ngoc-Duy, Chiang, Ya-Yu, Hardelauf, Heike, Waide, Sarah, Janasek, Dirk and West, Jonathan (2014) Preparation of neuronal co-cultures with single cell precision. Journal of Visualized Experiments, (87), e51389. (doi:10.3791/51389).

Record type: Article

Abstract

Microfluidic embodiments of the Campenot chamber have attracted great interest from the neuroscience community. These interconnected co-culture platforms can be used to
investigate a variety of questions, spanning developmental and functional neurobiology to infection and disease propagation. However, conventional systems require significant cellular inputs (many thousands per compartment), inadequate for studying low abundance cells, such as primary dopaminergic substantia nigra, spiral ganglia and Drosophilia melanogaster neurons, and impractical for high throughput experimentation. The dense cultures are also highly locally entangled, with few outgrowths (<10%) interconnecting the two cultures. In this paper straightforward microfluidic and patterning protocols are described which address these challenges: (i) a microfluidic single neuron arraying method, and (ii) a water masking method for plasma patterning biomaterial coatings to register neurons and promote outgrowth between compartments. Minimalistic neuronal co-cultures were prepared with high-level (>85%) inter-compartment connectivity and can be used for high throughput neurobiology experiments with single cell precision

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Published date: 20 May 2014
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 375895
URI: http://eprints.soton.ac.uk/id/eprint/375895
ISSN: 1940-087X
PURE UUID: 5fc6e2a0-0638-4a65-9055-f1669834bcb9
ORCID for Jonathan West: ORCID iD orcid.org/0000-0002-5709-6790

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Date deposited: 17 Apr 2015 14:03
Last modified: 15 Mar 2024 03:44

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Contributors

Author: Ngoc-Duy Dinh
Author: Ya-Yu Chiang
Author: Heike Hardelauf
Author: Sarah Waide
Author: Dirk Janasek
Author: Jonathan West ORCID iD

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