INVERTED MICROSCOPE WITH CONFOCAL ATTACHMENT

本提案用于购买Bio-Rad MicroRadiance共聚焦激光扫描显微镜系统，包括Nikon Eclipse TE 300倒置显微镜作为多用户共享仪器。 共聚焦显微镜将位于环境医学研究所并由其在PI的监督下维护。 业务和维护费用将从用户的年度捐款中回收，技术支助将从研究所的业务预算中提供。 在研究措施和治疗新生儿和成人急性肺部疾病的共同科学线索。 这些项目涉及组织损伤氧化剂的产生及其调节、细胞粘附分子功能、表面活性蛋白和脂质的遗传和病理生理调节、信号转导和细胞间通讯的研究。 这些项目将使用分离的大鼠和小鼠肺、培养的大鼠肺II型和平滑肌细胞以及其他可用的肺细胞系、人胎肺外植体、白细胞和大鼠脑血管。 这些研究通常在活细胞、厚外植体组织或离体肺中进行，需要使用荧光探针的共聚焦显微镜技术来可视化细胞生物学过程。 参与本提案的研究者开展需要对其研究领域相关分子进行细胞学定位和定量的研究，即：表面活性剂磷脂运输（Shuman），肺缺血性损伤中内皮功能障碍的评估（Fisher），PECAM-1的细胞分布和功能的评估（Albelda），测定表面活性蛋白SP-C加工的细胞和亚细胞位点（Beers），研究ICAM和硝基酪氨酸在CO中毒中的分布（Thom），评估C/EBP和proSP-C的分布（Feinstein），SP-B中间体和GPF运输的研究（Guttentag），气管平滑肌细胞中细胞骨架和信号转导蛋白的评价（Coburn），PECAM-1、GOX和过氧化氢酶的细胞分布的研究（Muzykantov），以及间隙连接蛋白连接蛋白43和26的共定位和运输的测定（Koval）。这些项目目前每年由NIH提供3，172，240美元的总费用支持，共聚焦显微镜的更大可用性将大大促进这些项目。

This proposal is for the purchase of a Bio-Rad MicroRadiance confocal laser scanning microscope system including a Nikon Eclipse TE300 inverted microscope as a multi-user shared instrument. The confocal microscope will be located in and maintained by the Institute for Environmental Medicine under the supervision of the PI. Operational and maintenance costs will be recovered from an annual contribution from the users, and technical support will be provided from the Institute's operational budget. The common scientific thread in the research measures, and treatment of acute lung diseases in the newborn and the adult. The projects involve study of generation of tissue- damaging oxidants and their regulation, cell adhesion molecule function, genetic and pathophysiologic regulation of surfactant proteins and lipids, signal transduction and cell-cell communication. The projects will use isolated rat and mouse lungs, cultured type II and smooth muscle cells from rat lungs and other available lung cell lines, human fetal lung explants, leukocytes and rat brain blood vessels. These studies, often in live cells, thick explant tissue, or isolated lung, require confocal microscopic techniques using fluorescence probes for visualization of cell biological processes. The investigators participating in this proposal carry out research requiring cytological localization and quantification of molecules pertinent to their fields of study, namely: surfactant phospholipid trafficking (Shuman), assessment of endothelial dysfunction in lung ischemic injury (Fisher), evaluation of cellular distribution and function of PECAM-1 (Albelda), determination of the cellular and subcellular sites of surfactant protein SP-C processing (Beers), study of distribution of ICAM and nitrotyrosine in CO-poisoning (Thom), assess distribution of C/EBP and proSP-C (Feinstein), study of trafficking of SP-B intermediates and GPF (Guttentag), evaluation of cytoskeletal and signal transduction proteins in tracheal smooth muscle cells (Coburn), study of cellular distribution of PECAM-1, GOX and catalase (Muzykantov), and determination of colocalization and trafficking of gap junction proteins connexins 43 and 26 (Koval). These projects which are currently supported by 3,172,240 dollars per annum in total costs from NIH will be greatly facilitated by the greater availability of a confocal microscope.