Intelligent Microtome Instrumentation

描述（由申请人提供）：在整个生物体中定位荧光标记细胞对基因治疗、干细胞治疗递送方法和药物研究的发展非常重要。本项目将开发一种智能切片机，用于识别全鼠荧光标记细胞，并将这些细胞收集成薄片。在第一阶段，将构建一个基于led的组织照明荧光成像的小型哺乳动物冷冻组。将对若干CCD摄像机进行评估，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和整只小鼠上进行测试。在这些样品中表达GFP的细胞将为第二阶段下一代仪器的开发提供数据。

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.