Macro Confocal Microscope System for Large-Scale Imaging in Basic and Translational Biology

 描述(由申请者提供)：我们请求资金购买最先进的商业共焦“宏镜”，配置为支持各种基本和翻译方法组合，需要在细胞、组织、器官和整个动物水平上对生物和疾病过程进行成像。该仪器是尼康AZ100-C2Si宏共焦显微镜系统，是专门为通过实时在宏观尺度上进行高分辨率荧光成像而设计的独一无二的仪器。尼康AZ-C2Si将在几个基本方面满足用户的需求。与标准共焦不同，该仪器具有大视场(对角线高达60 mm)和大工作距离(高达54 mm)。这些是超过其他共焦的数量级，使用户能够在不干扰光学的情况下对各种样品成像，这些样品是湿的或干的、干净的或原生的、非常厚的、在任何格式(例如，盘、盘和微流体环境)中安装或培养的。自动平滑变焦光学系统(8：1范围)将允许在同一样本中使用多个放大倍数进行延时实验，而不会有物体旋转进出光路。我们包括一个电动工作台，一个用于培养实验的气候室，以及一个四激光器配置，用于检测所有常用的染料组合和/或具有最高分辨率和成像灵敏度的荧光蛋白质变体。真光谱系统消除了荧光团之间的自发荧光和串扰，允许3D光学切片，并消除了焦外光线。在加州大学旧金山分校或市场上，没有其他工具拥有完全相同的功能和灵活性。正如提案中详细说明的那样，用户需要这些独特的功能来加速发现涉及大量活细胞和固定细胞、组织、器官和整个动物的项目。这项建议是基础、翻译和临床科学家的合作，他们在鸟类和哺乳动物中使用了各种模型系统。尼康AZ-C2Si将驻留在帕纳苏斯高地园区最近改建的实验室中，该实验室毗邻生物成像开发中心(BIDC)，并作为BIDC的一部分运营，BIDC是一个非常成功的核心设施，具有强大的机构支持 和加州大学旧金山分校的行政监督。尼康AZ-C2Si将首次在加州大学旧金山分校提供一套急需的特性和功能，这些特性和功能将导致组织工程和再生、干细胞分化、胚胎模式和出生缺陷、肿瘤进展和转移以及器官开发和修复等领域的新发现和重大研究进展。接入、培训和充值将由BIDC管理，BIDC在将创新技术转移到目前受现有工具技术限制的领域方面有着良好的记录。这一应用和工具将支持11名NIH资助的研究人员及其实验室，并促进至少23个NIH资助的R和U编码项目的进展。尼康AZ-C2Si也将提供给加州大学旧金山分校社区内的其他人。因此，这种共享仪器将解决用户未得到满足的需求，并加快加州大学旧金山分校的生物医学研究步伐。

 DESCRIPTION (provided by applicant): We request funds to purchase a state-of-the-art commercial confocal "macroscope" configured to support a diverse portfolio of basic and translational approaches that require imaging of biological and disease processes at the cellular, tissue, organ, and whole animal levels. The instrument, a Nikon AZ100-C2Si Macro Confocal Microscope System, is one-of-a-kind in being specially designed for high-resolution fluorescent imaging at a macroscopic scale in three dimensions through real time. The Nikon AZ-C2Si will meet the needs of its Users in several fundamental ways. Unlike standard confocals, this instrument has a large field of view (up to 60 mm diagonal) and large working distances (up to 54 mm). These are orders of magnitude beyond other confocals and will enable Users to image a wide range of samples that are wet or dry, cleared or native, very thick, and mounted or incubating in any format (e.g., dish, plate, and microfluidic environment) without interfering with the optics. An automated smooth zooming optical system (8:1 range) will allow for time-lapse experiments using multiple magnifications in the same sample without objectives rotating in and out of the light path. We include a motorized stage, a climate chamber for culture experiments, and a four-laser configuration for detecting all commonly used dye combinations and/or fluorescent protein variants with maximum resolution and imaging sensitivity. A true spectral system removes auto-fluorescence and cross talk between fluorophores, permits 3D optical sectioning, and eliminates out of focus light. No other instrument at UCSF or on the market has all the same capabilities and flexibility. As detailed in the proposal, Users need these unique features to accelerate discoveries on projects that involve large populations of live and fixed cells, tissues, organs, and whole animals. This proposal is a collaboration of basic, translational, and clinical scientists who use a variety of model systems in birds and mammals. The Nikon AZ-C2Si will reside in a recently remodeled laboratory on the Parnassus Heights Campus adjacent to, and operating as part of, the Biological Imaging Development Center (BIDC), which is a highly successful core facility that functions with strong institutional support and administrative oversight at UCSF. The Nikon AZ-C2Si will provide for the first time at UCSF a much-needed suite of features and functions that will lead to novel discoveries and dramatic research advances in areas such as tissue engineering and regeneration, stem cell differentiation, embryonic patterning and birth defects, tumor progression and metastasis, and organ development and repair. Access, training, and recharge will be managed by the BIDC, which has a proven track record of moving innovative technology into areas currently constrained by technical limitations of existing instruments. This application and instrument will support 11 NIH- funded investigators and their laboratories, and facilitate advances on at least 23 NIH-funded R- and U-coded projects. The Nikon AZ-C2Si will also be available to others within the UCSF community. Thus, this shared instrument will address unmet needs of its Users and accelerate the pace of biomedical research at UCSF.