A Confocal Microscope for Developmental and Cell Biological Analyses

用于发育研究的共聚焦显微镜- Skeath等人。该奖项支持购买激光扫描共聚焦显微镜，以跟踪活细胞，固定细胞，组织和发育生物体中的蛋白质表达。有了这种新型显微镜，华盛顿大学的研究人员将能够(1)通过厚的荧光标本切片薄的光学切片；（ii）同时观察这些标本中三种不同分子的亚细胞定位；（iii）跟踪多种蛋白在体内的表达和定位动态；（iv）将平行于光路的标本平面可视化；(v)获得所有标本的高分辨率三维视图。该显微镜将扩展圣路易斯华盛顿大学教师目前使用的实验技术。该提案详细介绍了新型共聚焦显微镜将产生重大影响的五个项目。这些项目包括：(1)分析控制果蝇胚胎中枢神经系统细胞多样性产生的发育和分子机制；（ii）了解细胞骨架和相关成分之间的相互作用如何驱动上皮细胞的形态变化，使用果蝇眼作为模型系统；（iii）追踪哺乳动物细胞系和小鼠胚胎中肌肉特异性基因的转录调控和功能；（iv）确定生殖细胞如何调节其进入减数分裂途径的决定及其进展；(v)阐明控制斑马鱼鳍再生和色素图案的分子机制。购买共聚焦显微镜的主要科学和教育目的是训练研究人员使用生命科学中最强大的显微镜之一，以便这些研究人员可以将共聚焦显微镜的功能与他们的研究结合起来。为了最大限度地利用显微镜，两名主要研究人员将与共聚焦制造商的一名代表一起，每月或根据需要进行培训。这些课程将确保使用显微镜的人员得到适当的培训。通过这种方式，pi希望让大学里任何感兴趣的人都能接触到共聚焦，包括博士后研究人员、研究生，希望还有本科生。

AWARD ABSTRACT - 9818013Confocal Microscope for Developmental Studies - Skeath et al.This award supports the purchase of a laser-scanning confocal microscope to follow protein expression in living as well as fixed cells, tissues, and developing organisms. With this new microscope, researchers at Washington University will be able (i) to slice thin optical sections through thick fluorescent specimens; (ii) to view simultaneously the sub-cellular localization of three different molecules within such specimens; (iii) to follow in vivo the expression and localization dynamics of multiple proteins; (iv) to visualize specimen planes parallel to the light path; and, (v) to obtain high resolution 3-D views of all specimens. The microscope will expand the experimental techniques presently employed by the faculty at Washington University in St. Louis. The proposal details five projects where the new confocal microscope will have a major impact. These projects include: (i) analyzing the developmental and molecular mechanisms that control the generation of cell diversity in the central nervous system of the Drosophila embryo; (ii) understanding how interactions between the cytoskeleton and related components drive morphological changes in epithelia using the Drosophila eye as a model system; (iii) tracking the transcriptional regulation and function of muscle-specific genes in mammalian cell lines and mouse embryos; (iv) determining how germ cells regulate their decision to enter, and their progression through, the meiotic pathway; and, (v) elucidating the molecular mechanisms that control fin regeneration and pigment patterning in the zebrafish, D. rerio. A primary scientific and educational goal of the acquisition of a confocal microscope is to train researchers in the use of one of the most powerful microscopes used in the life sciences, so that these researchers may integrate the capabilities of a confocal microscope with their research. To facilitate maximum utility of the microscope, two of the principal investigators together with a representative from the confocal manufacturer will carry out training sessions on a monthly basis or as demand dictates. These sessions will ensure proper training for those who use the microscope. In this way, the PIs hope to make the confocal accessible to any interested party at the university, including post-doctoral researchers as well as graduate and, hopefully, undergraduate students.