https://scholars.lib.ntu.edu.tw/handle/123456789/635633
標題: | Whole cell quenched flow analysis | 作者: | YA-YU CHIANG Haeri, Sina Gizewski, Carsten Stewart, Joanna D. Ehrhard, Peter Shrimpton, John Janasek, Dirk West, Jonathan |
公開日期: | 3-十二月-2013 | 卷: | 85 | 期: | 23 | 來源出版物: | Analytical Chemistry | 摘要: | 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. © 2013 American Chemical Society. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/635633 | ISSN: | 00032700 | DOI: | 10.1021/ac402881h |
顯示於: | 機械工程學系 |
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