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High-throughput optical screening of cellular mechanotransduction
Jonathan L Compton1, Justin C Luo2, Huan Ma1
1Department of Chemical Engineering and Materials Science, University of California, Irvine ; Laser Microbeam and Medical Program, Beckman Laser Institute, University of California, Irvine.
Nature Photonics
|October 14, 2014
Summary
We developed a novel optical platform using laser-generated micro-cavitation bubbles to rapidly screen molecules affecting cellular mechanotransduction without microfluidics.
Area of Science:
- Biophysics
- Cell Biology
- Optical Engineering
Background:
- Cellular mechanotransduction is crucial for physiological processes.
- Existing methods for studying mechanotransduction are often low-throughput.
- High-throughput screening of molecules affecting mechanotransduction is needed.
Purpose of the Study:
- To develop a rapid, high-throughput optical platform for screening molecules impacting cellular mechanotransduction.
- To investigate the use of laser-induced micro-cavitation bubbles (μCBs) to initiate mechanotransduction.
- To assess the platform's capability in real-time imaging and modulation analysis of cellular responses.
Main Methods:
- Utilizing single laser-microbeam generated micro-cavitation bubbles (μCBs) to induce hydrodynamic shear stress (microTsunami).
- Applying the microTsunami to stimulate adherent cells, specifically primary human endothelial cells.
- Imaging calcium (Ca2+) signaling as a readout of mechanotransduction and assessing modulation by exogenous molecules.
Main Results:
- Successfully initiated mechanotransduction in adherent cells using laser-generated μCBs and microTsunamis.
- Observed Ca2+ release consistent with G-protein-coupled receptor stimulation.
- Demonstrated dose-dependent inhibition of microTsunami-induced Ca2+ signaling by an exogenous inhibitor.
- Screened small molecules affecting cellular mechanotransduction in 96-well plates using imaging cytometry.
Conclusions:
- The developed optical platform enables rapid, high-throughput screening of molecules affecting cellular mechanotransduction.
- Laser-induced microTsunamis provide a microfluidics-free method to initiate and study cellular mechanotransduction.
- The platform allows for real-time assessment and modulation analysis of mechanosignaling pathways.
