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.
11560-11567
Chiang, Ya-Yu
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Haeri, Sina
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Gizewski, Carsten
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Stewart, Joanna D.
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Ehrhard, Peter
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Shrimpton, John
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Janasek, Dirk
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West, Jonathan
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3 December 2013
Chiang, Ya-Yu
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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
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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), .
(doi:10.1021/ac402881h).
(PMID:24295019)
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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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
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Date deposited: 03 Jan 2014 16:36
Last modified: 15 Mar 2024 03:43
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Contributors
Author:
Ya-Yu Chiang
Author:
Sina Haeri
Author:
Carsten Gizewski
Author:
Joanna D. Stewart
Author:
Peter Ehrhard
Author:
Dirk Janasek
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