Lightsheet Microscope for the UAB High-Resolution Imaging Facility

摘要 我们要求资金购买一个共享的光片荧光显微镜（LSFM），这是所需的多样化 一组NIH资助的研究人员在伯明翰的亚拉巴马大学（UAB），以提高我们的能力， 对光学透明和活体样本进行高分辨率、大体积成像。这个新的国家- 艺术显微镜将支持在UAB高分辨率成像设施（HRIF）-一个既定的显微镜 核心，将监督操作，培训和服务。LSFM可以让我们对完整的组织器官 类器官、胚胎和生物体的固定和清除以及活的制剂。UAB目前缺乏一个系统 用于高分辨率大体积成像。LSFM采用了一种新的配置，有利于快速高分辨率 完整组织和生物体的3D/4D成像，无需物理切片。考虑到科学的需要， 我们的用户群，我们选择蔡司Lightsheet 7显微镜作为最佳选择，以满足我们的要求， 在一定范围的样本大小、折射率和 决议。八个NIH资助的主要用户迫切需要LSFM推进他们的研究计划。LSFM 将为我们的研究小组提供几个主要的技术优势。首先，LSFM在高分辨率下采集图像， 速度，因此从大样本中收集数据是快速和有效的，使我们能够对更大的样本进行成像 包括整个生物体和完整的清除组织。第二，LSFM 基本上减少了光漂白和光损伤。第三，样品不需要安装在 载玻片/盖玻片允许模型生物体和完整组织的更多生理成像。最后，样品可以 可以相对于成像平面旋转，允许理想的定向和多视图的收集。述光片 7将作为我们的研究小组在发育生物学的不同利益的协同路口， 肺病学和神经科学。八个主要用户将利用该仪器对活斑马鱼进行成像， 非洲爪蟾、类器官和骨器官培养物，以及清除的组织，包括肺、脑、小鼠胚胎， 肌肉. Lightsheet 7将位于已建立的大学范围内的HRIF显微镜核心， 促进来自UAB各地的不同调查人员的访问。有强有力的制度支持， 这台显微镜与大学的大量贡献和长期承诺，以支持 核心对空间、人员和行政服务的需求。私家侦探是荧光方面的专家 在开发和应用新技术研究细胞生物学方面拥有20多年的经验 做了10年的显微镜中心主任所要求的显微镜将由 具有5年与HRIF用户合作和培训HRIF用户的经验的科学家，使用各种活样本和固定样本 在荧光显微镜中。进一步的支持将由一个7人监督委员会和一个 显微镜、组织清理和活体样本专家。蔡司Lightsheet 7将带来至关重要的成像 这些能力将推动我们NIH赞助的研究。

Abstract We request funds to purchase a shared light sheet fluorescent microscope (LSFM) that is required for a diverse group of NIH-funded investigators at the University of Alabama at Birmingham (UAB) to advance our abilities to perform High Resolution, large volume imaging of both optically cleared and live samples. This new state-of-the- art microscope will be supported in the UAB High Resolution Imaging Facility (HRIF) - an established microscopy core that will oversee operation, training, and service. LSFM will allow us to image intact tissues, organs, organoids, embryos, and organisms in both fixed and cleared and live preparations. UAB currently lacks a system for high resolution large-volume imaging. LSFM employs a novel configuration that facilitates fast high resolution 3D/4D imaging of intact tissues and organisms without physical sectioning. Considering the scientific needs of our user group, we selected the Zeiss Lightsheet 7 microscope as the best choice to meet our requirements for state-of-the-art live and optically cleared tissue imaging at a range of sample sizes, refractive indices, and resolutions. Eight NIH-funded major users critically require LSFM to advance their research programs. LSFM will provide our research group with several major technical advantages. First, LSFM acquires images at high speeds, and thus data collection from large samples is rapid and efficient, allowing us to image far larger samples than we are currently capable of, including whole organisms and intact cleared tissues. Second, LSFM substantially reduces photobleaching and photodamage. Third, samples do not need to be mounted on a slide/coverslip allowing more physiological imaging of model organisms and intact tissues. Finally, samples can be rotated relative to the imaging plane, allowing ideal orientation and collection of multiple views. The Lightsheet 7 will serve as a synergistic junction for our research group with diverse interests in developmental biology, pulmonology, and neuroscience. The eight major users will utilize the instrument to image live zebrafish, xenopus, organoids, and bone organ cultures, and cleared tissues including lung, brain, mouse embryos, and muscle. The Lightsheet 7 will be located in the established and university-wide HRIF microscopy core, thereby facilitating access by a diverse group of investigators from across UAB. There is strong institutional support for this microscope with substantial contributions from the university and a long-standing commitment to supporting the needs of the core for space, personnel, and administrative services. The PI is an expert in fluorescence microscopy with over 20 years of experience in developing and applying novel techniques to study cell biology and 10 years as a microscopy core director. The requested microscope will be supported day-to-day by a scientist with 5 years of experience working with and training HRIF users with a variety of live and fixed samples in fluorescence microscopy. Further support will be provided by a 7-person oversight committee and a team of microscopy, tissue clearing and live sample experts. The Zeiss Lightsheet 7 will bring critically important imaging capabilities that will advance our NIH-sponsored research.