SBIR-STTR Award

This Small Business Innovation Research (SBIR) Phase I project is aimed at developing an integrated system to stabilize a conventional high-end, inverted optical microscope to the precision required to routinely achieve the imaging power of emerging super-resolution (SR) microscopy techniques. These recent methods, enabled by particular photo-physical properties of certain fluorescent probes, have circumvented the diffraction limit normally associated with light microscopy and pushed it into realms thus far only achievable with electron microscopy. These methods put very high demands on the stability of the microscopy system. Conventional microscopes fail to meet these demands, and sample drift occurs relative to the image detector that can destroy the SR capabilities of the imaging system if this drift is not compensated. This project will integrate a 3-axis piezo-driven nanopostioning microscope stage with a closed-loop feedback system using data generated from an EMCCD camera, which will also serve as the image detector for the system. Fluorescent fiduciary references sparsely distributed within the sample will serve as anchor points from which to assess sample drift, and produce compensatory movement using the nanopositioning stage to stabilize the sample relative to the image detector in all 3 dimensions, in real-time. The broader impact/commercial potential of this project lies in making localization-based SR imaging methods routinely useful to working biologists. This means making them technically straightforward to implement, and economically accessible. Presently, there is no single source for the components required to stabilize a conventional microscope and make it routinely useful for SR microscopy. Scientists hoping to implement these very broadly applicable techniques are left with the daunting task of assembling a working system from its many individual components, and then getting these components to work together seamlessly as required. Our goal is to develop and then commercialize a fully-integrated microscope stabilization and imaging system based on a nanopositioning stage, an EMCCD camera, and software enabling real-time image-based feedback control of the sample?s position. It will be developed with localization-based SR microscopy in mind, but will be useful for any imaging experiment that requires long-term sample stability and image acquisition, such as extended live-cell imaging. This system will greatly simplify the technical challenges faced by biologists wanting to utilize SR microscopy as an experimental tool, enabling them to convert and extend their current inverted microscopes into potentially a large number of SR-capable imaging systems

This Small Business Innovation Research (SBIR) Phase II project will develop and commercialize an integrated system to actively stabilize an optical microscope to the precision required by today's cutting-edge imaging methods. Microscopy in the biological sciences is undergoing radical advancement on several fronts. "Super-Resolution" (SR) techniques circumvent the diffraction limit on resolution once thought to be insurmountable, and promise the ability to image the structures and processes of cell biology at the molecular level. This will usher in profound advancements in the understanding of the inner-workings of the cell. However, significant interrelated barriers remain in the path towards widespread use of SR techniques: (1) they are technically challenging and (2) expensive to implement; and (3) they place physical demands on the microscope platform it was not designed to meet. Foremost of these demands is that SR methods require control over the movement of the biological sample and the stability of the microscope system with nanometer precision. This commercialized integrated system is designed specifically to address these issues and remove these barriers. It uses a 3-axis, piezo-driven nanopositioning stage to control sample motion and actively maintains the stability of the system using the image as the reference point for this stability. The broader impact/commercial potential of this project lies in making SR methods routinely useful to working biologists. These "game-changing" tools will advance our understanding of the molecular bases of disease pathologies, and enable far more exacting methods aimed at their treatments. The new insights will range from those in molecular virology and the development of safer and more effective vaccines, to the molecular mechanisms of neuronal signaling and learning and memory. In fact, it is hard to imagine an area of cell biology that will not be impacted by these emerging SR techniques. One of the pioneers of these methods has likened them to the Hubble telescope: they enable people to see things they simply could not see before. This analogy goes further: there is only one Hubble telescope, and currently very few SR-capable imaging systems, due to both the technical and economic barriers to their routine use. And while SR methods expose the physical limitations of microscopes in an acute manner, their stability and image acquisition requirements are not unique. Thus, this commercial system will be much more broadly useful: it will also enable focal-stability and molecular tracking at the nanometer-scale for any long-term imaging experiment.