A Confocal Microscope for Developmental and Cell Biological Analyses

该奖项支持购买激光扫描共聚焦显微镜，以跟踪活的以及固定的细胞、组织和发育中的生物体中的蛋白质表达。使用这种新型显微镜，华盛顿大学的研究人员将能够(I)通过厚厚的荧光样本进行薄层光学切片；(Ii)同时观察此类样本中三种不同分子的亚细胞定位；(Iii)在体内跟踪多种蛋白质的表达和定位动态；(Iv)观察平行于光路的样本平面；以及(V)获得所有样本的高分辨率三维视图。该显微镜将扩展目前圣路易斯华盛顿大学教职员工使用的实验技术。该提案详细说明了新的共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；(2)了解细胞骨架和相关成分之间的相互作用如何推动上皮细胞的形态变化；(3)在哺乳动物细胞系和小鼠胚胎中跟踪肌肉特异基因的转录调控和功能；(4)确定生殖细胞如何通过减数分裂途径调节它们进入减数分裂的决定以及它们的进程；以及(5)阐明控制斑马鱼FIN再生和色素模式的分子机制。获得共焦显微镜的一个主要科学和教育目标是培训研究人员使用生命科学中使用的最强大的显微镜之一，以便这些研究人员可以将共焦显微镜的能力与他们的研究相结合。为了最大限度地发挥显微镜的效用，两名首席调查员将与来自共焦制造商的一名代表一起，每月或根据需求举办培训班。这些课程将确保对使用显微镜的人进行适当的培训。通过这种方式，PI希望让大学里任何感兴趣的人都可以访问共焦，包括博士后研究人员以及研究生，希望是本科生。

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.