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)，这是多样化所需的 美国国立卫生研究院资助的阿拉巴马大学伯明翰分校(UAB)的一组研究人员，以提高我们的能力 对光学清晰度和实时样品执行高分辨率、大体积成像。这个新的最新状态- ART显微镜将在UAB高分辨率成像设备(HRIF)中得到支持-一种已建立的显微镜 将监督运营、培训和服务的核心。LSFM将使我们能够对完整的组织、器官、 固定的、清除的和活的制剂中的有机体、胚胎和生物体。UAB目前缺乏一套系统 用于高分辨率的大体积成像。LSFM采用了一种新的配置，可实现快速高分辨率 没有物理切片的完整组织和生物体的3D/4D成像。考虑到科学上的需求 在我们的用户群中，我们选择了蔡司LightSheet 7显微镜作为满足以下要求的最佳选择 最先进的实时和光学透明组织成像，具有不同的样本大小、折射率和 决心。NIH资助的八个主要用户迫切需要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，使我们能够成像更大的样本 超过我们目前的能力，包括完整的生物体和完整的清除组织。第二，LSFM 大大减少光漂白和光损伤。第三，样本不需要安装在 幻灯片/盖片允许对模型生物和完整组织进行更多的生理成像。最后，样本可以 可以相对于成像平面旋转，从而可以理想地定位和收集多个视图。灯光片 7将成为我们在发育生物学领域有不同兴趣的研究小组的协同结合点， 肺病学和神经科学。八个主要用户将利用该仪器对活斑马鱼进行成像， 非洲爪哇、有机体和骨器官培养，以及清除的组织，包括肺、脑、小鼠胚胎和 肌肉。灯片7将位于已建立的大学范围内的HRIF显微镜核心中，从而 促进来自UAB各地的不同调查小组的访问。有强有力的制度支持 这台显微镜由大学做出了巨大贡献，并长期致力于支持 核心对空间、人员和行政服务的需求。PI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有20多年经验的显微镜 做了10年的显微镜核心主管。所要求的显微镜将由一名 有5年使用和培训HRIF用户的经验的科学家，使用各种活的和固定的样本 在荧光显微镜下。进一步的支持将由一个7人监督委员会和一个由 显微镜、组织清洗和活体样本专家。蔡司灯片7将带来至关重要的成像 这些能力将推进我们由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.