title: High-throughput optical screening of cellular mechanotransduction.
abstract: We introduce an optical platform for rapid, high-throughput screening of exogenous molecules that affect cellular mechanotransduction. Our method initiates mechanotransduction in adherent cells using single laser-microbeam generated micro-cavitation bubbles (μCBs) without requiring flow chambers or microfluidics. These μCBs expose adherent cells to a microTsunami, a transient microscale burst of hydrodynamic shear stress, which stimulates cells over areas approaching 1mm2. We demonstrate microTsunami-initiated mechanosignalling in primary human endothelial cells. This observed signalling is consistent with G-protein-coupled receptor stimulation resulting in Ca2+release by the endoplasmic reticulum. Moreover, we demonstrate the dose-dependent modulation of microTsunami-induced Ca2+signalling by introducing a known inhibitor to this pathway. The imaging of Ca2+signalling, and its modulation by exogenous molecules, demonstrates the capacity to initiate and assess cellular mechanosignalling in real-time. We utilize this capability to screen the effects of a set of small molecules on cellular mechanotransduction in 96-well plates using standard imaging cytometry.
authors.affiliation: Department of Chemical Engineering and Materials Science, University of California, Irvine ; Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Compton
authors.affiliation: Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine ; Department of Biomedical Engineering, University of California, Irvine.
authors.forename: Justin C
authors.initials: JC
authors.lastname: Luo
authors.affiliation: Department of Chemical Engineering and Materials Science, University of California, Irvine ; Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine.
authors.forename: Huan
authors.initials: H
authors.lastname: Ma
authors.affiliation: Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine ; Department of Biomedical Engineering, University of California, Irvine ; Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California, Irvine.
authors.forename: Elliot
authors.initials: E
authors.lastname: Botvinick
authors.affiliation: Department of Chemical Engineering and Materials Science, University of California, Irvine ; Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine ; Department of Biomedical Engineering, University of California, Irvine.
authors.forename: Vasan
authors.initials: V
authors.lastname: Venugopalan