Intelligent Microtome Instrumentation

智能切片机仪器

• 批准号: 7154878
• 负责人: Jeffrey J Kelly
• 金额: $ 10万
• 依托单位: BARLOW SCIENTIFIC, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7154878
• 关键词: artificial intelligence; bioimaging /biomedical imaging; biomedical equipment; biomedical equipment development; computer program /software; computer system hardware; digital imaging; fluorescence microscopy; genetically modified animals; green fluorescent proteins; laboratory mouse; molecular /cellular imaging; muscle; muscular dystrophy; phantom model; sectioning; three dimensional imaging /topography

DESCRIPTION (provided by applicant): Locating fluorescently-tagged cells in whole organisms is important to the development of gene therapy, stem cell therapy delivery methods, and for drug research. This project will develop an intelligent microtome to identify fluorescently labeled cells in whole mice and to collect those cells in thin sections. During Phase I, a small-mammal cryomicrotome with LED-based tissue illumination for fluorescence imaging will be constructed. Several CCD cameras will be evaluated to establish the spatial resolution and gray-scale resolution needed to provide necessary fluorescence-imaging capabilities for the instrument. The instrument will be tested on isolated muscle, tissue homogenates containing green fluorescent protein (GFP) labeled cells, and on whole mice. The cells expressing GFP in these samples will provide data for the development of next-generation instrumentation during Phase II. Phase II will add capabilities to use the fluorescence image data to intelligently acquire thin sections only when fluorescent cells are detected. Fluorescence images acquired during sectioning will provide morphological guides to identify and isolate the same cells in mounted sections by laser-capture microdissection. Automated thin-tissue acquisition will be an added capability and will be integrated with intelligent fluorescence analysis to produce a completely automated instrument. The instrument will allow processing of whole animals to obtain thin sections for laser dissection and for biochemical and/or PCR post processing of select tissue regions.

描述（由申请人提供）：在整个生物体中定位荧光标记细胞对于基因治疗、干细胞治疗递送方法和药物研究的发展非常重要。该项目将开发一种智能切片机来识别整只小鼠中的荧光标记细胞并将这些细胞收集成薄片。在第一阶段，将建造一个小型哺乳动物冷冻切片机，配备用于荧光成像的 LED 组织照明。将评估多个 CCD 相机，以确定为仪器提供必要的荧光成像功能所需的空间分辨率和灰度分辨率。该仪器将在分离的肌肉、含有绿色荧光蛋白（GFP）标记细胞的组织匀浆以及整只小鼠上进行测试。这些样品中表达 GFP 的细胞将为第二阶段的下一代仪器的开发提供数据。 第二阶段将增加仅在检测到荧光细胞时使用荧光图像数据智能采集薄片的功能。切片过程中获取的荧光图像将为通过激光捕获显微切割识别和分离固定切片中的相同细胞提供形态学指导。自动薄组织采集将是一项附加功能，并将与智能荧光分析集成以生产完全自动化的仪器。该仪器将允许对整个动物进行处理，以获得用于激光解剖的薄片以及对选定组织区域进行生化和/或 PCR 后处理。