A Confocal Fluorescence Microscope for the Study of the Cytoskeleton

这是一个购买扫描激光共聚焦荧光显微镜的小组建议书。该显微镜将是一个新的共享荧光显微镜设备的核心，该设备将成为研究人员研究项目的重点，并成为面向本科生的新实验室课程的关键元素。六名研究人员中的每一位都研究发育系统中细胞骨架的各个方面，重点是阐明细胞结构重塑和活动细胞器的组织。所有项目的目标都是利用高分辨率荧光显微镜来确定细胞骨架组件在发育过程中是如何组织和重新组织的。由于研究人员使用的实验系统(植物细胞、海胆卵和胚胎、果蝇胚胎、水蚤神经系统纤毛虫原虫)的复杂性、厚度和光学不透明性，传统的免疫荧光往往不足以满足研究的质量。共聚焦荧光显微镜的应用解决了这个问题；研究人员在初步的共聚焦显微镜工作中清楚地证明了这一点。遗憾的是，设在其他部门、可能用于这些研究的现有仪器严重超额认购，而且费用高昂。因此，尽管研究人员在优秀的模型系统和高度特定的工具中提出了令人着迷的问题，但他们受到设备缺乏的严重限制。这组研究人员的大小适合共用一台显微镜。而显微镜将被证明是这些研究小组之间持续密切互动的自然刺激。该提案的第二个重要特点是，共焦显微镜将用于许多学生的教育和培训。目前，在6个实验室中，有18名研究生和17名本科生，这35名学生包括14名女性，4名非裔美国人和4名亚裔美国人。这里描述的项目将由这些学生执行，因此他们将能够直接使用最先进的设备。第二种令人兴奋的方式是将这些设备用作一门新的本科实验室课程的核心，该课程由六名研究人员教授。本课程将使本科生能够设计、实施和讨论使用常规和共聚焦免疫荧光显微镜开发系统中细胞骨架的微型项目。

This is a group proposal to purchase a scanning laser confocal fluorescence microscope. The microscope will be the centerpiece of a new Shared Facility for Fluorescence Microscopy which will serve as the focus of the investigators' research projects and as the key element in a new laboratory course for undergraduates. Each of the six investigators studies aspects of the cytoskeleton in a developing system, focussing on the elucidation of cytoarchitectural remodeling and the organization of motile organelles. The goal of all of the projects is to utilize high resolution fluorescence microscopy to determine how the cytoskeletal components organize and re-organize during development. Because of the complexity, thickness, and optical opacity of the experimental systems used by the investigators (plant cells, sea urchin eggs and embryos, Drosophila embryos, leech nervous system ciliated protozoans), conventional immunofluorescence is often of insufficient quality for the studies. The problem is solved by the application of confocal fluorescence microscopy; this is clearly demonstrated in the preliminary confocal microscopy work by the investigators. Unfortunately, the existing instruments, located in other departments and which might be used for these studies, are heavily oversubscribed and costly. Thus, although the investigators are asking fascinating questions in excellent model systems and with highly specific tools, they are severely limited by a lack of equipment. The group of investigators is of the ideal size to share a microscope. And the microscope will prove to be a natural stimulus for the continued close interactions among these research groups. A second important feature of the proposal is that the confocal microscope will be utilized in the education and training of many students. Currently, among the six laboratories, there are 18 graduate students and 17 undergraduate students; these 35 students include 14 women, 4 African Americans, and 4 Asian Ameri cans. The projects described here will be performed by these students, and they will therefore be able to work directly with state-of-the-art equipment. A second exciting way the equipment will be utilized is as the centerpiece in a new undergraduate laboratory course, taught by the six investigators. The course will enable undergraduates to design, implement, and discuss mini-projects focussed on the cytoskeleton in developing systems using conventional and confocal immunofluorescence microscopy.