Technology Development

技术开发

• 批准号: 8157417
• 负责人: Michael Emmert-Buck
• 金额: $ 43.52万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8157417
• 关键词: Architecture; Biological Sciences; Cooperative Research and Development Agreement; Development; Dissection; Elements; Gel; Histology; Image; Individual; Knowledge; Membrane; Methods; Microdissection; Molecular; Molecular Analysis; Phenotype; Process; Proteins; RNA; Scanning; Series; Standardization; System; Techniques; Tissue Microarray; Tissues; Transcript; Work; base; gel electrophoresis; indexing; molecular pathology; technology development

We are currently working on developing two new technologies; expression microdissection (xMD) and layered expression scanning (LES). Based on immuno-targeting, xMD converts microdissection from an operator-dependent to an operator-independent mode, thus eliminating the need to laboriously procure cells. When fully developed xMD likely will offer several advantages over current methods, including; a) increased dissection rate (several orders of magnitude), b) increased dissection precision (subcellular), c) removal of variance among individual operators, permitting standardization of the dissection process, and d) elimination of targeting difficulties due to poor image quality of non-coverslipped (non-index-matched) histology sections used in current dissection systems. Layered expression scanning (LES) is a new technology co-developed by the Pathogenetics Unit and 20/20 GeneSystems, Inc. (http://www.2020gene.com/) through a Cooperative Research and Development Agreement (CRADA). The method utilizes a layered array of membranes for high-throughput molecular analysis and can be applied to a variety of life science platforms, including tissue sections, cells in culture, electrophoresis gels, multi-well plates, and tissue arrays. The technique is performed by passing the sample through a series of membrane layers while maintaining the original two-dimensional architecture. Each membrane measures a different molecular species, thus multiple RNA transcripts or proteins in each sample element (e.g., various cellular phenotypes in a tissue section, bands on a gel, individual wells of a microtiter plate) can be simultaneously analyzed. The method is conceptually simple, requires no moving parts, and can be used as an open or closed format.

我们目前正在开发两项新技术；表达显微切割（xMD）和分层表达扫描（LES）。基于免疫靶向，xMD 将显微切割从依赖操作员的模式转变为不依赖操作员的模式，从而消除了费力获取细胞的需要。当完全开发时，xMD 可能会比当前方法提供多种优势，包括： a) 提高解剖率（几个数量级），b) 提高​​解剖精度（亚细胞），c) 消除个体操作员之间的差异，允许解剖过程标准化，以及 d) 消除由于图像质量差而导致的瞄准困难当前解剖系统中使用的非盖玻片（非索引匹配）组织学切片。分层表达扫描 (LES) 是由病理遗传学部门和 20/20 GeneSystems, Inc. (http://www.2020gene.com/) 通过合作研究与开发协议 (CRADA) 共同开发的一项新技术。该方法利用分层膜阵列进行高通量分子分析，可应用于各种生命科学平台，包括组织切片、培养细胞、电泳凝胶、多孔板和组织阵列。该技术是通过使样品穿过一系列膜层来执行的，同时保持原始的二维结构。每个膜测量不同的分子种类，因此可以同时分析每个样品元素中的多个 RNA 转录物或蛋白质（例如，组织切片中的各种细胞表型、凝胶上的条带、微量滴定板的各个孔）。该方法概念简单，不需要移动部件，并且可以用作开放或封闭格式。