Single Cell Molecular Biology Through Electron Microscopy

通过电子显微镜进行单细胞分子生物学

• 批准号: 8902567
• 负责人: Michael Beer
• 金额: $ 2.5万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-15 至 1992-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8902567&HistoricalAwards=false
• 关键词: Single; Cell; Molecular; Biology; Through

This award provides funds to test the use of scanning electron microscopy for detection of minute quantities of the types of macromolecules (DNA, RNA, protein) found in all living organisms. The proposed test will use basic blotting techniques that involve adsorption of the macromolecule to a nitrocellulose or nylon membrane which is then exposed to antibodies or other, highly specific detector macromolecules that are themselves labelled with gold particles. Even a single particle of gold is readily visualized in the microscope. In principle, the successful development of the proposed technique would permit detection of a small number, possibly as few as one or two, molecules of the target macromolecule in a single cell. The use of general, but highly specific, readily quantitated assays for particular proteins, RNAs and DNAs has been crucial to the astounding progress of molecular biology in the last two decades. As now applied, most of these assays are applied to material extracted from large numbers of cells either grown in culture or present in a tissue, organ or even an entire organism. As yet, only a few, highly specialized tests are able to detect the minute quantities of such molecules present in single cells. The extension of more general techniques to permit analysis of low levels of unique molecules in single cells appears crucial to progress in understanding the control of development of well- differentiated multicellular organisms. Although not without risk, the approach to be undertaken has the potential of providing such an extension.

该奖项提供资金，以测试使用扫描电子 显微镜检测微量的类型， 大分子（DNA，RNA，蛋白质）存在于所有生物体中。 的 拟议的测试将使用基本的印迹技术，包括 将大分子吸附到硝化纤维素或尼龙膜上 然后将其暴露于抗体或其他高度特异性的检测器， 这些大分子本身被金颗粒标记。 甚至 在显微镜中容易看到单个金颗粒。 在 原则上，所提出的技术的成功发展将 允许检测少量，可能少至一个或两个， 在单个细胞中的靶大分子的分子。 使用通用但高度特异性、易于定量的检测方法 对于特定的蛋白质来说，RNA和DNA是至关重要的。 分子生物学在过去二十年里取得了惊人的进步。 作为 现在应用，大多数这些测定适用于提取的材料， 从大量的细胞，无论是在培养中生长，或存在于一个 组织、器官或甚至整个生物体。 到目前为止，只有少数，高度 专门的测试能够检测到微量的此类物质 存在于单个细胞中的分子。 更一般的延伸 技术允许分析低水平的独特分子， 单细胞似乎是理解控制的关键 分化良好的多细胞生物体的发展。 虽然并非没有风险，但所采取的办法具有 提供这种扩展的可能性。