Whole cell quenched flow analysis

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

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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

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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