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22 February 2013 Nonlinear structured illumination microscopy with surface plasmon resonance enhanced stimulated emission depletion
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Abstract
Nonlinear structured illumination microscopy (SIM) allows full-field imaging at resolutions <100 nm. Two nonlinear effects, excitation saturation (SSIM) and the photo-switching of protein had been applied to nonlinear SIM. We report a new SIM technique which utilizes the nonlinearity of STED effect. Resolution and signal noise ratio simulation shows that STED-SIM may serve as a better alternative to SSIM and SIM with photo-switchable protein. SIM requires a strong nonlinear effect in a large area. We use Surface Plasmon Resonant to enhance of evanescence field near a dielectric-metal-dielectric interface. An 8 times STED effect enhancement is achieved on an optimized glass-silver-glass-water planar structure. We further use the interference of two SPR-enhanced STED fields propagating at opposite direction to generate a 1D structured STED field. Combined with a uniform excitation field, the structure STED field allows full field total internal reflection imaging with an enhanced resolution along the structured dimension. Less than 50 nm resolution is demonstrated. A STED-SIM microscope with 2D structured STED field is under development. Future research will apply the microscope to superresolution imaging of membrane resident or near membrane structure at super-resolution in live cells.
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Han Zhang, Ming Zhao, and Leilei Peng "Nonlinear structured illumination microscopy with surface plasmon resonance enhanced stimulated emission depletion", Proc. SPIE 8590, Single Molecule Spectroscopy and Superresolution Imaging VI, 859011 (22 February 2013); https://doi.org/10.1117/12.2005043
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