CONFOCAL MICROSCOPE

共焦显微镜

• 批准号: 6051611
• 负责人: DIANA L KUNZE
• 金额: $ 19.11万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6051611
• 关键词: bioimaging /biomedical imaging; biomedical equipment purchase; confocal scanning microscopy

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.

这笔赠款要求提供资金来升级 Noran Odyssey XL 共焦显微镜，该显微镜目前是 Rammelkamp 研究所共享设施的一部分。该机器在进入市场后不久就被购买了。自最初购买 Odyssey XL 以来，图像采集和分析的能力得到了增强。升级（Noran OZ）将提供重要且所需的附加激光器和软件，具有解卷积能力，并最终提供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。下面列出的主要研究人员通常在两个领域使用本机器。一项主要用途是使用免疫细胞化学试剂、荧光追踪示踪剂和 GFP 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质。组织包括表达特定离子通道构建体的培养细胞系（HEK 细胞、L 细胞）、培养细胞（成纤维细胞、内皮 sf9 细胞）、原代培养物（神经元、肌细胞、上皮细胞、甲状旁腺）和组织切片（脑、心脏、肾） 。第二个主要用途是钙、膜电位和其他指标等染料，以评估这些离子在细胞功能中的作用。在这种方法中，我们利用该技术的时间分辨率以及空间分辨率。这些测量与电生理测量相结合。它们已用于研究内皮细胞和其他细胞响应激动剂的局部钙流入，以及动脉压力感受器末端电压敏感染料变化的研究。