An Enhanced Combinatorial Super-Resolution Microscopy Platform

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RTI funding will be used to enhance the spatial, temporal, and spectral resolution of an exceptionally capable super-resolution combinatorial microscopy platform that combines the sub-nanometer 3-D resolution of scanning probe microscopy with super-resolution fluorescence microscopy. This latter technique (and its variants) was recently awarded the 2014 Nobel Prize. The platform to be upgraded is unique in Canada with capabilities lacking in commercial systems. Comprising a 15 year old scanning probe microscope (SPM), a home-built, self-funded 4-laser dual-camera total internal reflection microscope, and a 4-laser confocal microscope, this platform is critical to multiple successful inter-disciplinary, -faculty, and -university collaborations, broadly supporting projects in structural biology, biophysics, and engineering. The present proposal seeks funds to replace the current SPM with a state-of-the-art system that has enhanced scan performance, significantly reduced noise, and higher sensitivity and resolution. This replacement is critically required as the current SPM is no longer supported by the vendor, and has become increasingly unreliable. Components cannot be sourced as they are no longer being manufactured leaving eBay and other auction sites as the only alternatives for parts. We are also seeking funding for a 561 nm laser to be integrated into the existing optical train (405 nm / 473 nm / 532 nm / 637 nm laser lines). This line will excite many of the key fluorophores for super-resolution imaging. The open-beam design of our optical train allows for ready control over phase, alignment, polarization, and beam shape, attributes that are not as easily manipulated within commercial systems.