BIOMEDICAL RESEARCH SUPPORT

这一修订后的提案要求为激光扫描共聚焦激光成像系统提供资金。 显微镜（LSCM）和适当的附件。 申请人是九个 来自解剖学，生长和发育， 药物化学、药学、生理学和口腔医学（涵盖 牙科、医学和药学院）。 的研究项目 调查人员需要详细的三维信息， 分子在细胞外和细胞内空间的定位。 项目描述涉及LSCM的应用，包括整体 组织样品以及细胞和亚细胞事件。 具体而言是 将探索以下重要领域：表皮屏障 发展和增强经皮转运、迁移和 早期胚胎细胞的侵袭活性，肿瘤细胞侵袭基底 膜和间质基质;受体介导的内吞作用和 抗原肽的摄取、生物化学和生物学表征 转化生长因子（TGF-α和TGF-β）， 极化上皮细胞，神经元的调节机制， 烟碱乙酰胆碱受体，以及 通过培养中的细胞的寡核苷酸。 对于每一个概述的研究计划， LSCM分析和量化空间分布的能力 分子穿过细胞膜，在细胞内和细胞周围，是至关重要的。 这 传统的信息技术无法实现复杂信息的质量， 免疫荧光显微镜，也不能准确地评估它使用 电镜 大量增加多项研究 方向将导致，因此，从共焦的可用性， 显微镜此时 虽然加州大学旧金山分校有这样的仪器， 目前调查小组的出入受到很大限制， 变得越来越困难。 药学院 政府已承诺空间的位置，一个新的设备， 这个应用程序是成功的。 因此，我们认为， 我们对这种共享设备有着重要的需求， 我们的机构没有其他选择，而且一个强大的 部门和学校对长期运作的承诺， LSCM的维护已经到位。 申请的仪器是BioRad- 600激光扫描共聚焦显微镜。 整个系统将包括 多个相互连接的组件：共焦成像系统，标准 荧光显微镜、视频打印机和光盘 存储.

This revised proposal requests funding for a laser scanning confocal microscope (LSCM) and appropriate accessories. The applicants are nine faculty members from the Departments of Anatomy, Growth and Development, Pharmaceutical Chemistry, Pharmacy, Physiology, and Stomatology (spanning the Schools of Dentistry, Medicine and Pharmacy). The research programs of the investigators require detailed information on the three-dimensional localization of molecules in extracellular and intracellular space. Projects are described which involve applications of LSCM both to whole tissue samples and to cellular and subcellular events. Specifically, the following areas of significance will be explored: epidermal barrier development and enhancement of percutaneous transport, migratory and invasive activity of early embryonic cells, tumor cell invasion of basement membrane and interstitial matrices; receptor-mediated endocytosis and the uptake of antigenic peptides, biochemical and biological characterization of transforming growth factors (TGF-alpha and TGF-beta), protein traffic in polarized epithelial cells, the regulatory mechanism(s) of neuronal nicotinic acetylcholine receptors, and the uptake and processing of oligonucleotides by cells in culture. For each research program outlined, the ability of the LSCM to analyze and quantify the spatial distribution of molecules through membranes, and within and around cells, is crucial. This quality of sophisticated information cannot be achieved with conventional immunofluorescence microscopy, nor can it be accurately assessed using electron microscopy. Considerable augmentation of multiple research directions will result, therefore, from the availability of a confocal microscope at this time. Although there are such instruments at UCSF, access for the present investigative team is very restricted and is becoming progressively more difficult. The School of Pharmacy administration has committed space for the location of a new apparatus in the event that this application is successful. We believe, therefore, that we have an important need for this shared equipment, that feasible alternatives are not available at our institution, and that a strong Departmental and School commitment to the long-term operation and maintenance of the LSCM is in place. The instrument requested is a BioRad- 600 laser-scanning confocal microscope. The overall system will consist of several interconnected components: confocal imaging system, standard epifluorescence microscope, video printer, and optical disk for mass storage.