Real-time Volumetric IR Absorption Nanoscope for in-situ Study of Plant/Microbe Interaction and Metabolism, 32a

Plant biologists currently lack viable, non-destructive, high-resolution microscopic imaging tools, necessary for investigating the real-time metabolism and biochemistry of plant and microbial cells. Existing methods are typically invasive and destructive, and require large and heavy instrumentation or access to national user facilities. Such instruments cannot be readily deployed in either public or private research settings, and are thus not accessible to the larger scientific community. This severely limits the understanding of complex plant and microbial sub-cellular processes, critical to the development of bioenergy and biofuels. The proposed effort will develop a turn- key, portable, living cell volumetric spectro-microscopic imager that will provide in vivo, time-resolved three dimensional infrared absorption (chemical) and visible (structural) imaging, with nanoscale lateral and axial spatial resolutions in both modalities. The nanoscope is based on a novel technique where localized transient photothermal changes in a specimen, induced by chemically-specific infrared absorption, are probed with exquisite sensitivity using visible-light digital holographic microscopy. The instrument’s compact form factor and user extensibility will enable its wide-scale deployment in the scientific community and provide researchers with the ability to visualize and understand complex inter- and intracellular biochemistries and interactions in living plant and microbial cells in the time domain. The Phase I effort will establish the feasibility of the proposed approach. Specifically, a system model describing all aspects of the proposed imager will be developed. A breadboard version of the imager will be assembled using primarily in-house components to demonstrate the capabilities of the proposed approach by generating proof of concept time-resolved volumetric infrared images of relevant plant and microbial cells. Finally, a conceptual design for a fully assembled prototype to be developed in the Phase II program, will be generated. The proposed imager will introduce a currently non- existent capability to the scientific community; an infrared spectroscopic imaging capability for plant biology research in government, commercial and academic sectors, directly benefiting and facilitating the advancement of bioenergy and biofuel development. The imager is also applicable to the wider array of life science, materials research and biomedical research. Applications include development of therapeutics and drug discovery. The imager is also applicable to biowarfare defense and will facilitate development of biological warfare agent countermeasures.