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Label-free visualization and quantification of single cell activity using metal-cald waveguide (MCWG) based microscopy

Thomas Söllradl, CTR

Evanescent-field based methods such as surface plasmon resonance (SPR) have been used very effectively for label-free imaging of microscopic biological material in close proximity to a sensing surface. However, the shallow probing depth of SPR (typically less than ~200 nm) can be problematic when imaging relatively thick biological objects such as cells or bacteria. In this paper, we demonstrate how metal-clad waveguides (MCWG) can be used to achieve deeper probing depth compared to SPR while maintaining good imaging spatial resolution. Comparative numerical simulations of imaging spatial resolution versus probing depth are shown for a  number of common SPR, long-range SPR, and MCWG configurations, demonstrating that MCWG offer the best compromise between resolution and depth for imaging thick biological objects. We demonstrate the potential of this approach by monitoring intracellular activity following the activation of apoptosis in individual cells exposed to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and by visualizing and quantifying extracellular changes in endothelial cell layer integrity following the activation of the proteinase-activated receptor 1 (PAR1) by thrombin.

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