MOLECULAR DYNAMICS MULTIPROBE CONFOCAL LASER MICROSCOPE

该提案要求提供资金购买一台分子动力学2001 共聚焦激光扫描显微镜。这台仪器将服务于 贝勒医学院一些教职员工的研究需求 研究工作的重点是活细胞的动力学，三个- 亚细胞结构的空间排列，以及三个- 完整脑内神经元等细胞的三维重建 切片。我们有相当数量的调查人员，他们的工作包括 活细胞显微镜从分子水平延伸到 组织中细胞之间的相互作用。我们在BCM的调查人员是 熟悉数字成像的优势和局限性 荧光显微镜。专业知识的多样性包括光学 显微镜、分子生物学、光谱学、图像处理、分析 化学、显微注射、免疫化学、细胞生物学、分子 图形学、结构和计算生物学。除 提供卓越的空间分辨率优势 波长共聚焦显微镜的主观判读 细胞现象，这项技术的应用是由 BCM调查人员还有另外两个共同主题。第一个是需要 关于亚细胞的三维组织的信息 结构和组织中随时间变化的响应 特定的微扰。第二个问题是需要量化细胞 特征和/或流程：尺寸、出现频率、 摄取或细胞器如何运动--属性可以由 一个数字。研究从分子水平延伸到 组织中细胞之间的相互作用已经达到了由 较老的显微镜方法的主观性以及在 手动处理，创建数据库所需的足够信息 采用统计学方法进行分析。总体而言，我们有经验和 建立合作以利用最先进的多波长 用于收集和分析新型信息的共焦仪器 使用统计标准，从而提高生产率和价值 我们正在进行的研究。尽管我们较老的显微镜方法和 定量数字荧光成像的最新发展已经 对我们来说很好，我们的研究计划已经到了一个点 如果有最先进的光学仪器，进展更快 显微镜是可用的。在过去的几年里，设施致力于 光学显微镜和电子显微镜已经发展成为一种 德克萨斯医学中心、莱斯大学和 休斯顿大学。尽管有许多成熟的和富有成效的 协作，我们需要利用最近在 光学显微镜。很难从厚厚的制剂中获得图像 数字格式，用来处理这些问题的计算方法是 不能令人满意。更高分辨率的图像将对 推进和整合我们在显微镜方面的项目。共焦 请在此申请仪器，作为来自 凯克计算生物学中心代表着一个必不可少的 我们的超微结构和数字显微镜中心的成分，并将 作为贝勒研究社区的共享资源， 德克萨斯大学医学院、莱斯大学和德克萨斯大学 休斯敦。

This proposal requests funds to purchase a Molecular Dynamics 2001 Confocal Laser Scanning microscope. This instrument will serve the research needs of a number of faculty of Baylor College of Medicine whose research work focuses on the dynamics of living cells, the three- dimensional arrangement of subcellular structures, and the three- dimensional reconstruction of cells such as neurons in intact brain slices. We have a critical mass of investigators whose work involves microscopy of living cells extending from the molecular level to interactions between cells in tissues. Our investigators at BCM are conversant with the advantages and limitations of digital imaging fluorescence microscopy. The diversity in expertise includes optical microscopy, molecular biology, spectroscopy, image processing, analytical chemistry, microinjection, immunochemistry, cell biology, molecular graphics and structural and computational biology. In addition to the benefits of superior spatial resolution that are provided by multi wavelength confocal microscopy of the subjective interpretation of cellular phenomena, the applications of this technology envisioned by the BCM investigators have two other themes in common. The first is a need for information about the three-dimensional organization of subcellular structures and the time dependent change in their organization in response to specific perturbants. The second is a need to quantify cellular features and/or processes: dimensions, frequency of occurrence, rate of uptake or how an organelle moves -- attributes that can be represented by a number. Investigations extending from the molecular level to interactions between cells in tissues have reached the limits imposed by the subjectivity of older microscopic methods and by the difficulty in processing manually, sufficient information to create databases for analysis by statistical methods. Collectively, we have the experience and collaborations established to utilize state-of-the-art multiwavelength confocal instrumentation to collect and analyze new kinds of information using statistical criteria, thereby increasing the productivity and value of our ongoing research. Although our older microscopic methods and the more recent development of quantitative digital fluorescence imaging has served us well, we have reached a point where our research programs would progress faster if state-of-the- art instrumentation for optical microscopy were available. Over the past several years facilities devoted to light and electron microscopy have been developed that serve as a research resource for the Texas Medical Center, Rice University, and the University of Houston. Despite many established and productive collaborations, we need to take advantage of recent developments in optical microscopy. Images from thick preparations are difficult to obtain in digital format and the computational methods used to treat these are not satisfactory. Higher resolution images will be critical for the advancement and integration of our programs in microscopy. The confocal instrumentation requested here as part of the $100,000 matching funds from the Keck Center for Computational Biology represents an essential ingredient in our ultrastructural and digital microscopy center and would serve as a shared resource to the research community at Baylor, the University of Texas Medical School, Rice University, and the University of Houston.