SYSTEM FOR QUANTITATIVE 3D CONFOCAL LIGHT MICROSCOPY

从功能破坏的角度来理解疾病的能力， 健康的细胞通常受到我们对功能的了解的限制。 正常组织的结构。 这些知识是慢慢获得的，因为 研究细胞结构的工具要么使用起来非常乏味， （电子显微镜）或在正常厚度的细胞中无效 （常规光学显微镜）。 该提案的目的是获得一个 用于研究细胞的3D功能结构的系统， 革命性的共焦光学显微镜新技术。 该系统 将由七名细胞生物学家组成的核心用户组使用， 神经生物学家，他们共同从事五个主要领域， research. 这些是1）c-癌基因在细胞生长中的作用， 分化; 2）心脏和骨骼发育的早期步骤 肌肉; 3）神经组织subserving处理的视觉 肌球蛋白分子的结构特征 负责骨骼肌原纤维粗丝的组装， 5)参与收缩性血管形成的途径和分子 肌肉和非肌肉细胞中的结构。 这一核心用户群体研究的主题多种多样。 但我们 分享需要一种快速的方法来研究细胞结构在0.1到 1.0 mu分辨率范围，一种不会因组织而变得无用的方法 厚度在0.1-0.5 mm范围内。我们共同探索了使用 激光扫描共聚焦显微镜，发现这种新开发的 这项技术为我们的研究开辟了令人兴奋的新可能性。

The ability to understand disease in terms of the disruption of function in healthy cells is frequently limited by our knowledge of the functional architecture of normal tissue. This knowledge is gained slowly because tools for studying cell structure have been either very tedious to use (electron microscopy) or ineffective in cells of normal thickness (conventional light microscopy). The aim of this proposal is to acquire a system for studying the 3D functional architecture of cells based on the revolutionary new techniques of confocal light microscopy. This system will be utilized by a core user group of seven cell biologists and neurobiologists who collectively are engaged in five major areas of research. These are 1) role of c-oncogenes in cell growth and differentiation; 2) early steps in the development of heart and skeletal muscle; 3) neural organization subserving the processing of visual information in the retina; 4) structural features of the myosin molecule responsible for the assembly of thick filaments of skeletal myofibrils and 5) pathways and molecules involved in the formation of contractile structures in muscle and non-muscle cells. The subjects studied by this core user group are diverse. However, we share the need for a rapid way of studying cell architecture in the 0.1 to 1.0 mu resolution range, a way that is not rendered useless by tissue thickness in the range 0.1-0.5 mm. We have jointly explored the use of laser scanning confocal microscopy and found that this newly developed technique opens up exciting new possibilities for our research.