MRI: Development: Extreme Life Volumetric Imaging System (ELVIS)

该项目的目标是开发一种独特的多模态体积显微镜，能够对比传统荧光显微镜采样的样品体积大约100倍的样品体积进行瞬时成像，其空间分辨率足以检测大多数细菌和古菌。该仪器将首次联合收割机结合两种模式：数字全息显微镜（DHM）（包括振幅[明场]和定量相位成像）和荧光光场显微镜（FLFM）。Co-PI已经将每种技术分别作为实验室的研究仪器;这将是第一次将具有所有所述功能的显微镜用于研究。将建造的仪器将被包装用于水生环境中的现场使用。它将在实验室中使用，并提供给研究人员。PSU正在对其研究基础设施，计划和长期前景进行重大改造，其中包括对STEM的兴趣日益增长和博士课程的重大扩展。该工具将使研究和培训跨越学术水平，从高中到博士，并将成为PSU的不断增长的研究概况和与其他机构和社区的参与的重要组成部分。PI有一个公共宣传和培训的既定模式;两个特别优秀的领域是（1）让本科生参与研究，特别强调招募女性和代表性不足的少数民族，包括英语学习者;（2）为STEM教师制定课程和培训计划。该工具将有助于这些努力以及新的努力，具体而言：在现有的初级/高级/研究生水平的细胞和分子生物物理学课程中增加实验室部分;创建新的高级顶点实地课程;参加美国宇航局的“空间前进”教师讲习班。第一个结合体积明场，相位和荧光成像系统，将由全息照相和光场显微镜结合而成。这将使瞬时体积成像，这在细胞，发育和有机体生物学以及物理科学（水文学，材料科学）中具有广泛的新兴和潜在的应用。传统的显微镜需要切片（共聚焦或多光子）太慢，无法捕捉大量的动力学事件：细菌游泳，心跳，血流，细胞内细胞器运动等等。如果可以在单个相机快照中记录扩展样本体积的空间信息，则可以减轻常规成像模式的限制。这种同步体积成像能力不仅消除了传统显微镜的精细聚焦要求，而且还大大提高了成像吞吐量。 在该项目中，PI将根据远程或极端环境的原位环境微生物学定制仪器。微尺度生物体仍然是一个被忽视的尺度，对其进行了有限的原位成像。ELVIS的具体目标区域包括公海、海冰、冰川冰和热液喷口。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.