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

该项目的目标是开发一种独特的多模式体积显微镜，能够对样品体积进行瞬时成像，样品体积大约是传统荧光显微镜样品体积的100倍，空间分辨率足以检测大多数细菌和古菌。该仪器将首次将两种模式结合在一起：数字全息显微镜(DHM)(包括幅度[亮场]和定量相位成像)和荧光光场显微镜(FLFM)。合作伙伴已将每项技术单独确定为其实验室中的研究仪器；这将是第一次提供具有所有所述能力的显微镜用于研究。将要建造的仪器将被打包，用于在水生环境中的野外使用。它将在实验室中使用，并向研究人员提供。PSU正在对其研究基础设施、计划和长期前景进行重大更新，其中包括对STEM的兴趣日益增长和博士项目的重大扩展。这一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日益增长的研究形象和与其他机构和社区接触的重要组成部分。公共宣传和培训具有既定的模式；两个特别突出的领域是：(1)吸引本科生参与研究，特别强调招聘妇女和代表不足的少数群体，包括英语学习者；(2)为STEM教师制定课程和培训方案。该仪器将帮助这些努力以及新的努力，特别是：在现有的初级/高级/研究生级别的细胞和分子生物物理学课程中增加一个实验室组件；创建一个新的高级Capstone野外课程；参加NASA“太空探索”教师工作坊。将通过耦合全息显微镜和光场显微镜创建第一个组合的体积亮场、相位和荧光成像系统。这将使瞬时体积成像成为可能，它在细胞、发育和生物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应用。需要切片(共聚焦或多光子)的传统显微镜太慢了，无法捕捉到大量的动力学事件：细菌游泳、心跳、血液流动、细胞内细胞器运动等。如果人们可以在单个相机快照中记录扩展样本体积的空间信息，则可以减轻传统成像方式的限制。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，而且显著提高了成像吞吐量。在这个项目中，PI将为偏远或极端环境的现场环境微生物学量身定做仪器。微米级的生物体仍然是一个被忽视的大小尺度，对其进行的原位成像有限。猫王的具体目标区域包括开阔的海洋、海冰、冰川冰和热液喷口。已经确定了来自不同机构的十几个感兴趣的用户。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.