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Whole cell quenched flow analysis

Whole cell quenched flow analysis
Whole cell quenched flow analysis
This paper describes a microfluidic quenched flow platform for the investigation of ligand-mediated cell surface processes with unprecedented temporal resolution. A roll-slip behavior caused by cell-wall-fluid coupling was documented and acts to minimize the compression and shear stresses experienced by the cell. This feature enables high-velocity (100-400 mm/s) operation without impacting the integrity of the cell membrane. In addition, rotation generates localized convection paths. This cell-driven micromixing effect causes the cell to become rapidly enveloped with ligands to saturate the surface receptors. High-speed imaging of the transport of a Janus particle and fictitious domain numerical simulations were used to predict millisecond-scale biochemical switching times. Dispersion in the incubation channel was characterized by microparticle image velocimetry and minimized by using a horizontal Hele-Shaw velocity profile in combination with vertical hydrodynamic focusing to achieve highly reproducible incubation times (CV = 3.6%). Microfluidic quenched flow was used to investigate the pY1131 autophosphorylation transition in the type I insulin-like growth factor receptor (IGF-1R). This predimerized receptor undergoes autophosphorylation within 100 ms of stimulation. Beyond this demonstration, the extreme temporal resolution can be used to gain new insights into the mechanisms underpinning a tremendous variety of important cell surface events.
0003-2700
11560-11567
Chiang, Ya-Yu
0ab4fe9e-9a3e-4594-a13c-73f68c13b8b3
Haeri, Sina
feee833a-70f4-4457-af71-7c696286074c
Gizewski, Carsten
04bb649a-d58a-47bf-8263-ceb3fb5716f6
Stewart, Joanna D.
e1ec9784-39cc-48ed-9f4f-2a05d25f2106
Ehrhard, Peter
c53d081f-b45d-4636-9232-a6636c47a789
Shrimpton, John
9cf82d2e-2f00-4ddf-bd19-9aff443784af
Janasek, Dirk
39a248d4-ac4a-4f07-9919-7bc1098cd715
West, Jonathan
f1c2e060-16c3-44c0-af70-242a1c58b968
Chiang, Ya-Yu
0ab4fe9e-9a3e-4594-a13c-73f68c13b8b3
Haeri, Sina
feee833a-70f4-4457-af71-7c696286074c
Gizewski, Carsten
04bb649a-d58a-47bf-8263-ceb3fb5716f6
Stewart, Joanna D.
e1ec9784-39cc-48ed-9f4f-2a05d25f2106
Ehrhard, Peter
c53d081f-b45d-4636-9232-a6636c47a789
Shrimpton, John
9cf82d2e-2f00-4ddf-bd19-9aff443784af
Janasek, Dirk
39a248d4-ac4a-4f07-9919-7bc1098cd715
West, Jonathan
f1c2e060-16c3-44c0-af70-242a1c58b968

Chiang, Ya-Yu, Haeri, Sina, Gizewski, Carsten, Stewart, Joanna D., Ehrhard, Peter, Shrimpton, John, Janasek, Dirk and West, Jonathan (2013) Whole cell quenched flow analysis. Analytical Chemistry, 85 (23), 11560-11567. (doi:10.1021/ac402881h). (PMID:24295019)

Record type: Article

Abstract

This paper describes a microfluidic quenched flow platform for the investigation of ligand-mediated cell surface processes with unprecedented temporal resolution. A roll-slip behavior caused by cell-wall-fluid coupling was documented and acts to minimize the compression and shear stresses experienced by the cell. This feature enables high-velocity (100-400 mm/s) operation without impacting the integrity of the cell membrane. In addition, rotation generates localized convection paths. This cell-driven micromixing effect causes the cell to become rapidly enveloped with ligands to saturate the surface receptors. High-speed imaging of the transport of a Janus particle and fictitious domain numerical simulations were used to predict millisecond-scale biochemical switching times. Dispersion in the incubation channel was characterized by microparticle image velocimetry and minimized by using a horizontal Hele-Shaw velocity profile in combination with vertical hydrodynamic focusing to achieve highly reproducible incubation times (CV = 3.6%). Microfluidic quenched flow was used to investigate the pY1131 autophosphorylation transition in the type I insulin-like growth factor receptor (IGF-1R). This predimerized receptor undergoes autophosphorylation within 100 ms of stimulation. Beyond this demonstration, the extreme temporal resolution can be used to gain new insights into the mechanisms underpinning a tremendous variety of important cell surface events.

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More information

e-pub ahead of print date: 12 November 2013
Published date: 3 December 2013
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 360763
URI: http://eprints.soton.ac.uk/id/eprint/360763
ISSN: 0003-2700
PURE UUID: 43cd374e-3463-458b-b924-0dd9cf8d066b
ORCID for Joanna D. Stewart: ORCID iD orcid.org/0000-0002-2608-1967
ORCID for Jonathan West: ORCID iD orcid.org/0000-0002-5709-6790

Catalogue record

Date deposited: 03 Jan 2014 16:36
Last modified: 21 Nov 2021 03:07

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Contributors

Author: Ya-Yu Chiang
Author: Sina Haeri
Author: Carsten Gizewski
Author: Joanna D. Stewart ORCID iD
Author: Peter Ehrhard
Author: John Shrimpton
Author: Dirk Janasek
Author: Jonathan West ORCID iD

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