SBIR Phase I: Widefield 3D Superresolution Microscopy Module

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This Small Business Innovation Research Program (SBIR) Phase I project investigates the feasibility of a ground-breaking three-dimensional (3D) optical microscope with single-molecule sensitivity suitable for live cell studies and nanometer scale sensing. Several methods have recently shown resolution well beyond the optical diffraction limit, which restricts imaging to features larger than about 200 and 500 nm in the transverse and axial dimensions respectively. To date, these super-resolution techniques have been demonstrated in specialized labs but new developments are required to reach the fundamental limits on a regular basis at the typical biology lab. Therefore, this project focuses on fundamental technological developments to address the upcoming need for pervasive 3D superresolution single-molecule capabitlities. The instrument is based on a joint design of the illumination, optical response, data collection approach, and reconstruction algorithms for fluorescence imaging. The instrument makes use of tailored point spread functions that enable the determination of the 3D position of emitters with great precision, leading to better resolution and depth of field than competing methods. Preliminary laboratory experiments have shown 3D capability with resolutions below 20nm, which corresponds to one order of magnitude enhancement over most optical microscopes presently in use. The broader impact/commercial potential of this project addresses a major opportunity in an expanding biological instrumentation market. If successful, it will enable 3D superresolution imaging capability to many biological laboratories currently limited by past technologies. The feasibility plan opens up opportunities for an accelerated commercialization path by implementing a cost-effective flexible solution compatible with existing microscopes. The extensive availability of superresolution imaging will impact several fields of science and engineering including 3D biophysical and biomedical optical imaging. During the project the company will generate synergistic relationships to rapidly transfer the technology into applications. The company has established partnerships for testing the instrument in significant biological problems that are expected to contribute to a better understanding of diseases. From a broader perspective, this project will increase scientific technical capabilities by providing nanoscale optical imaging for everyday biological research, thereby strengthening US presence in the worldwide optical microscopy industry.