Upright Light Microscope Workstation

正置光学显微镜工作站

• 批准号: 7047408
• 负责人: Ulrich Hopfer
• 金额: $ 18.71万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7047408
• 关键词: epithelium; lighting

DESCRIPTION (provided by applicant): This is a request for the upright epifluorescence light microscope Leica workstation DM 6000B with incubation chamber. The microscope set-up is needed for live cell studies of epithelial cell monolayers with dual-sided perfusion. The microscope is requested for a group of investigators concerned with investigating the dynamics and polarity of cell signaling, gene therapy, transport, and cellular organization of differentiated epithelial cells. It is well recognized by now that hormonal or growth factor regulation can occur not only from the blood, but also the luminal side in polarized cells. Similarly, flow on the luminal side affects cellular functions. Investigations of the dynamics and spatial organization of these regulatory processes is crucial for progress in understanding normal and pathological behavior of epithelial cells. Advances in cell culture methods have allowed us to establish in-vitro models for many different types of epithelia. To obtain highly differentiated cells and mimic the in-vivo situation, cells are grown on permeable support to form polarized, confluent, differentiated epithelial cell layers in which the dynamics of different cellular processes can be studied by fluorescence microscopy. Such epithelia are perfused with different solutions and/or rates on apical and basal side to mimic physiological conditions, i.e., nutrients, oxygen and certain regulatory agents on the basal (blood) side and other regulatory agents and flow conditions on the apical (luminal) side. In particular, we will study in live epithelia: flow dependence of receptor trafficking and signaling (renal proximal tubule cells); the intracellular fate of condensed polylysine/DNA complexes used for gene therapy (pulmonary epithelia); spatial regulation of EOF receptor signaling from wild type and polycystic kidney diseased mice (renal collecting duct cells), interactions of SI00 proteins during differentiation (epidermis), regulation of luminal pH (epithelia of the female reproductive tract), and the force required for cilia deflection (renal epithelial cells). These studies require a state-of-the-art upright epifluorescence light microscope workstation with temperature and atmospheric control for live cells and an optical chamber with dual perfusion and water immersion optics with high numerical aperture.

描述(申请人提供)：这是一个立式荧光显微镜徕卡工作站DM6000B带孵化室的要求。双面灌流的上皮细胞单层活细胞研究需要显微镜装置。显微镜是研究分化上皮细胞的细胞信号、基因治疗、运输和细胞组织的动力学和极性的研究人员所需要的。现在已经很好地认识到，激素或生长因子的调节不仅可以发生在血液中，而且还可以发生在极化细胞的管腔一侧。同样，管腔一侧的血流也会影响细胞功能。研究这些调控过程的动力学和空间组织对于理解上皮细胞的正常和病理行为是至关重要的。细胞培养方法的进步使我们能够建立许多不同类型的上皮细胞的体外模型。为了获得高度分化的细胞并模拟体内的情况，细胞在可渗透的载体上生长，形成极化的、融合的、分化的上皮细胞层，在这些细胞层中，可以用荧光显微镜研究不同细胞过程的动力学。这样的上皮细胞在顶端和基底端注入不同的溶液和/或速率，以模拟生理条件，即基底端(血液)的营养物质、氧气和某些调节剂，以及尖端(腔)端的其他调节剂和流动条件。特别是，我们将在活上皮细胞中研究：受体转运和信号传递的流动依赖性(肾近端小管细胞)；用于基因治疗的浓缩多聚赖氨酸/DNA复合体的细胞内命运(肺上皮)；野生型和多囊肾疾病小鼠(肾集合管细胞)EOF受体信号的空间调控；SI00蛋白在分化过程中的相互作用(表皮)；管腔pH的调节(女性生殖道上皮)；纤毛偏转所需的力(肾上皮细胞)。这些研究需要最先进的直立式荧光显微镜工作站，具有活细胞的温度和大气控制，以及具有大数值孔径的双重灌流和水浸光学的光学室。