Lightsheet Microscope for the UAB High-Resolution Imaging Facility

摘要 我们请求资金购买共享光片荧光显微镜 (LSFM)，这是多样化研究所需的 阿拉巴马大学伯明翰分校 (UAB) 的一组由 NIH 资助的研究人员致力于提高我们的能力 对光学透明样品和活体样品进行高分辨率、大体积成像。这种新的状态—— 艺术显微镜将得到 UAB 高分辨率成像设施 (HRIF) 的支持 - 一种成熟的显微镜 监督运营、培训和服务的核心。 LSFM 将使我们能够对完整的组织、器官、 固定、澄清和活体制剂中的类器官、胚胎和生物体。 UAB目前缺乏系统 用于高分辨率大体积成像。 LSFM 采用新颖的配置，有助于快速实现高分辨率 无需物理切片即可对完整组织和生物体进行 3D/4D 成像。考虑到科学的需要 我们的用户组，我们选择 Zeiss Lightsheet 7 显微镜作为满足我们要求的最佳选择 在各种样本大小、折射率和范围内进行最先进的实时光学透明组织成像 决议。 8 个由 NIH 资助的主要用户迫切需要 LSFM 来推进他们的研究项目。 LSFM 将为我们课题组提供几大技术优势。首先，LSFM在高处获取图像 速度，因此从大样本中收集数据是快速和高效的，使我们能够对更大的样本进行成像 比我们目前的能力更强，包括整个生物体和完整的透明组织。二、LSFM 显着减少光漂白和光损伤。第三，样品不需要安装在 载玻片/盖玻片允许对模型生物和完整组织进行更多的生理成像。最后，样品可以 相对于成像平面旋转，允许理想的方向和多个视图的收集。灯片 7 将作为我们研究小组在发育生物学方面具有不同兴趣的协同结合点， 肺病学和神经科学。八个主要用户将利用该仪器对活体斑马鱼进行成像， 爪蟾、类器官和骨器官培养物，以及清除的组织，包括肺、脑、小鼠胚胎和 肌肉。 Lightsheet 7 将位于已建立的全大学 HRIF 显微镜核心中，从而 方便来自 UAB 的不同研究人员群体进行访问。有强有力的制度支持 这台显微镜得到了大学的巨大贡献以及对支持的长期承诺 核心区对空间、人员和行政服务的需求。 PI是荧光方面的专家 显微镜技术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 并担任显微镜核心主任 10 年。所需的显微镜将由日常支持 拥有 5 年使用各种活体和固定样本与 HRIF 用户合作和培训经验的科学家 在荧光显微镜中。一个由 7 人组成的监督委员会和一个由 7 人组成的团队将提供进一步的支持 显微镜、组织透明化和活体样本专家。 Zeiss Lightsheet 7 将带来至关重要的成像 这些能力将推动我们 NIH 资助的研究。

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.