The objective of this project is to develop a unique multi-modal volumetric microscope capable of instantaneous imaging of sample volumes approximately 100-fold larger than those sampled by conventional fluorescence microscopy, with spatial resolution sufficient to detect most bacteria and archaea. This instrument will combine two modalities for the first time: Digital Holography Microscopy (DHM) (which includes amplitude [brightfield] and quantitative phase imaging) and Fluorescence Light Field Microscopy (FLFM). The Co-PIs have established each technology separately as research instruments in their laboratories; this will be the first time that a microscope with all of the described capabilities will be made available for research. The instrument that will be build will be packaged for field use in aquatic environments. It will be used in the lab, and also made available to researchers. PSU is undergoing a major renovation of its research infrastructure, plan, and long-term outlook, which includes a growing interest in STEM and a major expansion in PhD programs. This instrument will enable research and training across academic levels, from high school through PhD, and will be an important part of PSU's growing research profile and engagement with other institutions and the community. The PI has an established pattern of public outreach and training; two areas of particular excellence are (1) engaging undergraduates in research, with a particular emphasis on recruiting women and underrepresented minorities, including English language learners; and (2) developing curriculum and training programs for STEM teachers. This instrument will assist these efforts, as well as new efforts, specifically: addition of a laboratory component to the existing junior/senior/graduate-level Cellular and Molecular Biophysics course; creation of a new Senior Capstone field course; participation in the NASA "Spaceward Bound" teachers' workshops.
The first combined volumetric brightfield, phase, and fluorescence imaging system, will be created by coupling holographic and light field microscopy. This will enable instantaneous volumetric imaging, which has a wide variety of emerging and potential applications in cell, developmental, and organismal biology, as well as in the physical sciences (hydrology, materials science). Conventional microscopy requiring sectioning (confocal or multiphoton) is too slow to capture a large number of kinetic events: bacterial swimming, heartbeats, blood flow, intracellular organelle movement, and many more. The limitations of conventional imaging modalities could be mitigated if one could record the spatial information of an extended sample volume in a single camera snapshot. Such synchronous volumetric imaging capability not only removes the delicate focusing requirement of conventional microscopy, but also dramatically enhances the imaging throughput. In this project, the PI will tailor the instrument to in situ environmental microbiology of remote or extreme environments. Micron-scale organisms remain a neglected size scale for which limited in situ imaging has been performed. Specific target areas for ELVIS include the open ocean, sea ice, glacier ice, and hydrothermal vents. A dozen interested users from different institutions have been identified.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
