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的细胞对基因治疗、干细胞治疗给药方法的发展和药物研究都很重要。该项目将开发一种智能显微切割机，以识别整个小鼠的荧光标记细胞，并将这些细胞收集在薄片中。在第一阶段，将建造一个基于LED组织照明的小型哺乳动物冷冻微型机，用于荧光成像。将对几个CCD相机进行评估，以确定为该仪器提供必要的荧光成像能力所需的空间分辨率和灰度级分辨率。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个小鼠身上进行测试。这些样品中表达GFP的细胞将为第二阶段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获取薄片的功能。在切片过程中获得的荧光图像将提供形态指南，以通过激光捕获显微切割识别和分离安装在切片中的相同细胞。自动薄层组织采集将是一项额外的能力，并将与智能荧光分析相集成，以生产一种完全自动化的仪器。该仪器将允许对整个动物进行处理，以获得用于激光解剖和选定组织区域的生化和/或聚合酶链式反应后处理的薄片。