CAREER: Integrated Computational Optical Framework for Quantitative Space-Variant Imaging in Live-Cell Fluorescence Microscopy

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CAREER: Integrated computational optical framework for quantitative space-variant imaging in live-cell fluorescence microscopy This research is developing new computational methods to integrate with a novel optical non-scanning imaging system design to enable fast live-cell fluorescence microscopy imaging with high, isotropic 3D resolution. The PI has pioneered computational methods for DIC microscopy. This work addresses a key challenge in live-cell microscopy of accounting for aberrations due to ?thick? biological specimens. Refractive index (RI) mismatch and heterogeneity within thick samples produce image distortions that worsen with imaging depth in conventional microscopes. Commercially available deconvolution methods do not account for these distortions. To address this issue, the PI is utilizing computational Differential Interference Contrast (DIC) microscopy to develop new corrective methodologies and integrate computational development into a new system design using a computational optical sensing and imaging (COSI) system approach. This is a novel and innovative application of adaptive optics. Specific research is developing mathematical models that can accurately predict data acquired with a wide-field imaging system; using model-based data processing algorithms to estimate accurate fluorescence concentration in 3D images and developing a software package for the user community. Performance and utility of the new methods is being tested on cytoskeletal proteins. The broader impact activities are diverse and innovative. The PI is developing coursework that integrates imaging science and optical systems to complement research in computational imaging for microscopy and COSI system design. The PI is establishing cross-disciplinary participation in her courses, particularly among engineering, biological sciences, and pre-medicine majors. A interdisciplinary certificate program with concentration in Microscopy will result. The PI?s commitment to multidisciplinary education and outreach is preparing the next generation of biological researchers and engineers for work at the intersection of biology, computational science and engineering disciplines where significant advances are expected to further revolutionize science.