Single Cell Microinjection and Image Analysis in Developmental Biology

该提案是关于生物设施中心 发育生物学。 美国加州大学 建议设立及/或提升其单身人士设施。 细胞显微注射，图像分析，流式细胞术， 高压液相色谱法 单细胞 微注射设备将用于标记单个细胞 以便追踪它们的行踪和随后的命运 在不同的实验条件下;注射标记染料， 研究间隙、连接细胞通讯;注射 反义RNA或抗体干扰合成 或特定蛋白质的功能;并注射DNA， 遗传转化实验 两个显微镜工作站 具有图像处理和分析能力的 建立：一个用于荧光显微镜，一个用于 差示干涉显微术 荧光 显微镜将配备一个共聚焦激光扫描装置 可以直接收集数字化的荧光 图像具有虚拟消除的焦点外噪声以及 比传统成像更高的对比度和分辨率 技术.两个工作站都会将数据输入到 计算机化凝胶和放射自显影分析设备 使用视频密度计进行全面定性分析， 详细定量的软激光扫描密度计 问题研究 流式细胞仪设施将升级， 一个附加的激光管和一个计算机化的数据管理系统 系统 高压液相色谱系统还将 建立。

This proposal is for a Biological Facilities Center for Developmental Biology. The University of California proposes to establish and/or upgrade its facilities for single. cell microinjection, image analaysis, flow cytometry, and high.pressure liquid chromatography. The single.cell micro injection facility will be used to mark individual cells in order to follow their movements and subsequent fates under various experimental conditions; to inject marker dyes to investigate gap.junctional cell communication; to inject antisense RNA or antibodies to interfere with the synthesis orfunction of specific proteins; and to inject DNA for genetic transformation experiments. Two microscopy workstations with image processing and analysis capabilities will be established: one for fluorescence microscopy and one for differential interference contrast microscopy. The fluorescence microscope will be fitted with a confocal laser scanning device that allows the direct collection of digitized fluorescence images with virtual elimination of out.of.focus noise as well as greater contrast and resolution than conventional imaging techniques. Both workstations will feed data to a Microvax facility for computerized analysis of gels and autoradiograms using a video densitometer for overall qualitative analysis and a soft laser scanning densitometer for detailed quantitative studies. The flow cytometry facility will be upgraded by adding an additional laser tube and a computerized data management system. A high.pressure liquid chromatography system will also be established.