Instruments for Fluorescence and Video-Enhanced Microscopy

该提案要求为荧光和视频增强显微镜设备提供资金，包括用于检测低强度荧光的冷却电荷耦合器件（CCD）相机和成像系统，配备无限远校正和微分干涉对比光学器件的荧光显微镜，用于视频增强显微镜的图像处理系统和Newvicon相机，以及用于植物细胞的微操作器和压电微注射器。这是由来自四个系和两个学院的7名研究者（5名主要用户，Wendy F. Boss, Rebecca S. Boston, Mark Conkling, Dominique Robertson， Ron Sederoff和Ross Whetten，以及2名次要用户，Tim Petty和Kelly Tatchell）联合提出的建议。据我们所知，校园内没有其他制冷CCD摄像机，农业与生命科学学院或林业资源学院也没有类似的高分辨率视频增强显微镜设备。主要研究人员使用分子、遗传、生化和显微技术的组合来研究基本的细胞过程和信号转导。研究项目包括玉米的信号转导（Boss）、蛋白质运输和包装到储存囊泡（Boston）、根特异性基因表达和线虫抗性（Conkling）、病毒发病机制（Robertson和Petty）、细胞分裂和细胞周期控制（Tatchell）、木质素生物合成、火炬松的真菌抗病和遗传定位（Sederoff和Whetten）。在上述研究中，我们需要可视化活细胞内的荧光蛋白，监测蛋白质分布和细胞形态随时间的变化，并解析中期染色体上的荧光标记。我们的研究目前受到现有显微设备的灵敏度和分辨率的限制。植物系通过提供空间和技术支持对该设施作出了承诺。这些设备将由温蒂·博斯博士和达纳·莫克斯利先生管理，并将被安置在一个中心位置。所要求的设备将对我们正在进行的加强北卡罗莱纳州立大学细胞生物学的努力作出重大贡献。

This proposal requests funds for equipment for fluorescence and video-enhanced microscopy to include a cooled charged coupled device (CCD) camera and imaging system for detecting low intensity fluorescence, a fluorescence microscope equipped with infinity corrected and differential interference contrast optics, an image processing system and Newvicon camera for video-enhanced microscopy, and a micromanipulator and piezoelectric microinjector for use with plant cells. This is a joint proposal by 7 investigator (5 major users, Wendy F. Boss, Rebecca S. Boston, Mark Conkling, Dominique Robertson, Ron Sederoff and Ross Whetten, and 2 minor users, Tim Petty and Kelly Tatchell) from four departments and two colleges. To our knowledge there is no other cooled CCD camera on campus and there is no comparable equipment for high resolution video-enhanced microscopy in the College of Agriculture and Life Sciences or the College of Forestry Resources. The principle investigators study basic cellular processes and signal transduction using a combination of molecular, genetic, biochemical, and microscopic techniques. The research projects include signal transduction (Boss), protein trafficking and packaging to storage vesicles in maize (Boston), root-specific gene expression and nematode resistance (Conkling), viral pathogenesis (Robertson and Petty), cell division and cell cycle control (Tatchell), and lignin biosynthesis, fungal disease resistance and genetic mapping in loblolly pine (Sederoff and Whetten). For the above studies we need to visualize fluorescent proteins within living cells, to monitor changes in protein distribution and cellular morphology over time, and to resolve fluorescent markers on metaphase chromosomes. Our research is currently limited by the sensitivity and resolution of existing microscopic equipment. The Botany Department has made a commitment to the facility by providing space and technical support. The equipment will be under the s upervision of Dr. Wendy F. Boss and Mr. Dana Moxley, and it will be housed in a central location. The requested equipment will contribute significantly to our ongoing efforts to strengthen cell biology at North Carolina State University.