Single Cell Microinjection and Image Analysis in Developmental Biology

这是一个关于发育生物学的生物设施中心的提案。加州大学建议建立和/或升级其单一设施。细胞显微注射，图像分析，流式细胞术，和高。压力液相色谱法。单一。细胞微注射设备将用于标记单个细胞，以便在各种实验条件下跟踪其运动和随后的命运；注射标记染料观察间隙。连接细胞通讯；注射反义RNA或抗体以干扰特定蛋白质的合成或功能；并为基因转化实验注入DNA。将建立两个具有图像处理和分析能力的显微镜工作站：一个用于荧光显微镜，一个用于差干涉对比显微镜。荧光显微镜将配备一个共聚焦激光扫描装置，允许直接收集数字化荧光图像，虚拟消除焦外噪声，以及比传统成像技术更大的对比度和分辨率。这两个工作站将向Microvax设备提供数据，使用视频密度计进行全面定性分析，使用软激光扫描密度计进行详细定量研究，对凝胶和自射线照相进行计算机分析。流式细胞仪设备将通过增加一个额外的激光管和一个计算机数据管理系统进行升级。一个高。建立压力液相色谱系统。

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.