CONFOCAL MICROSCOPE

这笔拨款要求资金升级Noran Odyssey XL共聚焦显微镜，该显微镜目前是Rammelkamp研究所共享设施的一部分。这台机器刚投放市场不久就被买走了。自最初购买奥德赛XL以来，图像采集和分析的能力得到了增强。此次升级（Noran OZ）将提供具有反褶积功率的重要且必要的额外激光和软件，最终实现更高的空间和时间分辨率。培训、管理和会计程序已经在进行中。下面列出的主要调查人员目前使用的机器一般有两个方面。一个主要用途是利用免疫细胞化学试剂、荧光跟踪示踪剂和绿色荧光蛋白作为信号，以高分辨率识别和定位各种组织中的细胞内和细胞外蛋白。组织包括表达特定离子通道构建的培养细胞系（HEK细胞、L细胞）、培养细胞（成纤维细胞、内皮细胞）、原代培养（神经元细胞、肌细胞、上皮细胞、甲状旁腺细胞）和组织切片（脑、心、肾）。第二个主要用途是染料，如钙，膜电位和其他指标，以评估这些离子在细胞功能中的作用。在这种方法中，我们利用了该技术的时间分辨率和空间分辨率。这些测量与电生理测量相结合。它们已被用于研究内皮细胞和其他细胞对激动剂的局部钙内流反应，以及动脉压力感受器末端电压敏感染料变化的研究。

This grant requests funds to upgrade a Noran Odyssey XL confocal microscope presently part of the shared facilities in the Rammelkamp Research Institute. This machine was purchased shortly after its introduction into the market. The capacity for image acquisition and analysis has been enhanced since the original purchase of the Odyssey XL. The upgrade (Noran OZ) will provide important and needed additional laser and software with deconvolution power and, finally, higher spatial and time resolution. Training, management and accounting procedures are already in pace. The use of the present machine by the major investigators listed below has been in general two areas. One major use has been to identify and localized with high resolution the intracellular and extracellular proteins in a variety of tissues using immunocytochemical agents, fluorescent track tracers and GFP as signals. Tissue include cultured cell lines expressing particular ion channel constructs (HEK cells, L cells), cultured cells (fibroblasts, endothelial sf9 cells), primary cultures (neuronal, myocytes, epithelial, parathyroid), and tissue slices (brain, heart, kidney). The second major use has been dyes such as calcium, membrane potential and other indicators to assess the roles of these ions in cellular function. In this method we take advantage of the time resolution as will as spatial resolution of the technique. These measurements are combined with electrophysiological measurements. They have been used for studying localized calcium influx in endothelial and other cells in response to agonists and studies of voltage-sensitive dye changes in arterial baroreceptor terminals.