Intelligent Microtome Instrumentation

描述(申请人提供)：使用染料、荧光蛋白和量子点对细胞进行荧光标记，构成了现代生物学中许多强大分析方法的基础。在整个动物体内定位这些细胞对于研究细胞的传递、分化和基因表达是必要的。第二阶段的SBIR研究正在开发一种智能显微切割机，以提供在完整的器官或生物体内定位细胞并捕获包含目标细胞的薄片的方法。该仪器将对整个器官或小型哺乳动物进行连续切片，记录每一切片后组织的多波长荧光图像，实时分析荧光图案的变化，并自动从存在荧光标记细胞的区域收集薄片。这种紧凑的仪器将安装在典型的实验室工作台上。将在大学研究实验室建造仪器进行测试，研究基因治疗、干细胞行为、肺功能障碍和通过细菌转基因进行基因转移。在第一阶段对智能切片机的商业市场进行了利基分析。分析表明，人们对这种低成本的三维成像方法和选择性的自动薄片收集方法产生了广泛的兴趣。与公共卫生相关：正在设计新的生物医学方法，将DNA、RNA或干细胞输送到体内的目标位置，以治疗疾病。第二阶段SBIR提案中描述的研究将在第一阶段进行的原型开发研究的基础上制造智能显微镜。这些新仪器将在大学研究实验室进行评估，以确定其对整个研究动物进行切片和成像的能力，定位干细胞或表达DNA或RNA的细胞，并将这些细胞收集在薄片中进行进一步检查。这项研究将有助于回答“整个生物体内的哪些细胞会受到基因或干细胞疗法的影响？”以及“这些细胞在生化水平上发生了什么？”

DESCRIPTION (provided by applicant): Fluorescent tagging of cells using dyes, fluorescent proteins, and quantum dots forms the basis for many powerful analytical methods in modern biology. Locating these cells within entire animals is necessary to study cell delivery, differentiation and gene expression. The Phase II SBIR research is developing an intelligent microtome to provide the means to locate cells within an intact organ or organism and to capture thin sections containing target cells. This instrument will serially section whole organs or small mammals, record multiple wavelength fluorescence images from the tissue after removal of each section, analyze changes in fluorescent patterns in real time, and automatically collect thin sections from areas where fluorescently tagged cells are present. The compact instrument will mount on a typical laboratory bench. Instruments will be constructed for testing in university research laboratories studying gene therapy, stem cell behavior, pulmonary dysfunction, and gene transfer by bacterial transfection. A niche analysis of the commercial market for the intelligent microtome was performed during Phase I. The analysis indicated broad interest in this low-cost approach to three- dimensional imaging and the selective automated approach to collection of thin sections. PUBLIC HEALTH RELEVANCE: New biomedical methods are being designed to deliver DNA, RNA, or stem cells to targeted locations within the body to treat disease conditions. The research described in this Phase II SBIR proposal will manufacture intelligent microtomes, based upon prototype development studies conducted in Phase I. These new instruments will be evaluated in university research laboratories for their capability to section and image entire research animals, to locate stem cells or cells in which DNA or RNA is expressed, and to collect those cells in thin sections for further examination. This research will help the answer the questions "Which cells within the entire organism are affected by the gene or stem cell therapy?" and "What is going on at a biochemical level within these cells?"