Automated Slide Scanner with Brightfield and Fluorescence for Stereology, FISH and Brain Connectivity Mapping

摘要： 需要资金购买Aperio Versa 200自动幻灯片扫描仪，以引入三种 目前Gladstone的任何服务核心或 加州大学旧金山分校：i)以设计为基础的体视学(确保在分析中进行严格的实验设计 三维形态)ii)通过全幻灯片成像(WSI)和iii)定量组织FISH 大脑连接图(追踪整个大脑中特定回路的输入)。幻灯片扫描仪 将被安置在加州大学旧金山分校的格莱斯顿研究所组织学和光学显微镜中心， 格拉德斯通和加州大学旧金山分校的主要组织学服务提供商。仪器将会是 主要由8名调查人员使用，其中5人在Gladstone内部，3人来自其他地方 加州大学旧金山分校的部门。此请求中确定的主要用户包括NIH资助的研究人员 其项目侧重于影响人类健康的领域，包括心脏病、阿尔茨海默氏症 疾病、学习和记忆、再生医学、帕金森氏症和肥胖。此外 对于新的实验能力，该系统将使WSI的周转时间增加至少7- 与目前可用的收购方法相比，这是一种新的收购方式。Aperio Versa 200提供 全自动扫描多达200张1“x 3”幻灯片，放大倍数最高可达63倍。该系统包括 亮场和荧光摄像机，1.25x、5x/0.15、10x/0.32、20x/0.7、40x/0.85(带 校正项圈)和63x/1.25油目标，用于DAPI、GFP/FITC、Cy3、Cy5和 MCherry，12V 100W金属卤化物荧光照明器和自动加油系统 分辨率成像。这些组合的特点使它适合于广泛的实验。至 提供所需的具体实验能力，要求为软件模块提供资金 支持先进的体视学分析、多通道分割和定量(IHC或IFA)， 和组织鱼分析模块。结合起来，这一设备将极大地扩展核心能力 并允许快速、可重现且深入地探测大量 数量特征。预计此请求中确定的主要用户将占 大约80%的使用时间；在剩余的可访问用户时间内，幻灯片扫描仪 将可供其他领域的研究人员使用。幻灯片扫描仪将由公司的工作人员操作 Gladstone研究所的组织学和光学显微镜核心，由训练有素的用户提供。这台仪器 将允许核心提供基于幻灯片的多功能图像采集和分析模式 样本提供给格拉德斯通大学和加州大学旧金山分校的研究人员，从而显著扩大了该岩芯在 当地的研究界。

Abstract: Funds are requested to purchase the Aperio VERSA 200 automated slide scanner, to introduce three new imaging modalities currently lacking from any of the service cores at Gladstone or either of the UCSF campuses: i) Design-based stereology (to ensure rigorous experimental design in analysis of three-dimensional morphologies) ii) Quantitative tissue FISH by whole slide imaging (WSI) and iii) Brain Connectivity Mapping (to trace inputs to specific circuits in whole brains). The slide scanner would be housed at the Gladstone Institutes Histology and Light Microscopy Core at UCSF, the principal histological service provider for Gladstone and UCSF. The instrument will be predominantly used by 8 investigators, 5 of whom are within Gladstone and 3 from other departments at UCSF. The Major Users identified in this request include NIH -funded researchers whose projects are focused on areas affecting human health including heart disease, Alzheimer’s disease, learning and memory, regenerative medicine, Parkinson’s disease, and obesity. In addition to the new experimental capabilities, the system will increase turn-around-time for WSI by at least 7- fold, as compared to the currently available acquisition methods. The Aperio VERSA 200 provides fully automated scanning of up to 200 1” x 3” slides at up to 63x magnification. The system includes brightfield and fluorescence cameras, 1.25x, 5x / 0.15, 10x / 0.32, 20x / 0.7, 40x / 0.85 (with correction collar), and 63x / 1.25 oil objectives, filter cubes for DAPI, GFP/FITC, Cy3, Cy5, and mCherry, a 12V 100W metal halide fluorescence illuminator and an automated oiling system for high resolution imaging. These combined features make it suitable for a wide range of experiments. To provide the specific experimental capabilities required, funds are requested for software modules enabling advanced stereological analysis, multi-channel segmentation and quantitation (IHC or IFA), and tissue FISH analysis modules. Together, this equipment will greatly expand the core capabilities and allow rapid, reproducible, and deep-reaching probing of the data on a large number of quantitative features. It is predicted that the Major Users identified in this request will account for approximately 80 % of the usage time; during the remaining Accessible User Time the slide scanner will be available for other area researchers. The slide scanner will be operated by the staff of the Histology and Light Microscopy Core at the Gladstone Institute and by trained users. This instrument will allow the core to offer versatile image acquisition and analysis modalities from slide-based samples to researchers at Gladstone and UCSF, thus significantly expanding this core’s utility for the local research community.