Whole cell quenched flow analysis

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

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)11560–11567
Number of pages8
JournalAnalytical Chemistry
Volume85
Issue number23
Early online date31 Oct 2013
DOIs
Publication statusPublished - 3 Dec 2013

Fingerprint

Microfluidics
Ligands
IGF Type 1 Receptor
Insulin
Cell membranes
Velocity measurement
Shear stress
Demonstrations
Hydrodynamics
Imaging techniques
Computer simulation
Convection
Intercellular Signaling Peptides and Proteins

Keywords

  • microfluidic quenched flow
  • temporal resolution
  • autophosphorylation

Cite this

Chiang, Y-Y., Haeri, S., Gizewski, C., Stewart, J. D., Ehrhard, P., Shrimpton, J., ... West, J. (2013). Whole cell quenched flow analysis. Analytical Chemistry, 85(23), 11560–11567. https://doi.org/10.1021/ac402881h
Chiang, Ya-Yu ; Haeri, Sina ; Gizewski, Carsten ; Stewart, Joanna D. ; Ehrhard, Peter ; Shrimpton, John ; Janasek, Dirk ; West, Jonathan. / Whole cell quenched flow analysis. In: Analytical Chemistry. 2013 ; Vol. 85, No. 23. pp. 11560–11567.
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Chiang, Y-Y, Haeri, S, Gizewski, C, Stewart, JD, Ehrhard, P, Shrimpton, J, Janasek, D & West, J 2013, 'Whole cell quenched flow analysis', Analytical Chemistry, vol. 85, no. 23, pp. 11560–11567. https://doi.org/10.1021/ac402881h

Whole cell quenched flow analysis. / Chiang, Ya-Yu; Haeri, Sina; Gizewski, Carsten; Stewart, Joanna D.; Ehrhard, Peter ; Shrimpton, John; Janasek, Dirk; West, Jonathan.

In: Analytical Chemistry, Vol. 85, No. 23, 03.12.2013, p. 11560–11567.

Research output: Contribution to journalArticle

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AU - Chiang, Ya-Yu

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Chiang Y-Y, Haeri S, Gizewski C, Stewart JD, Ehrhard P, Shrimpton J et al. Whole cell quenched flow analysis. Analytical Chemistry. 2013 Dec 3;85(23):11560–11567. https://doi.org/10.1021/ac402881h