Laser Scanning Confocal Microscope

本提案要求购买并操作一台Bio-Rad共聚焦显微镜和一台Nikon rE 2000 U倒置显微镜，以加强Kosair儿童医院研究所（KCHRI）快速发展的研究项目。KCHRI自2000年成立以来，这个小组已经招募了8名教师和大量的研究科学家，博士后研究员和学生，共有40名研究人员。他们中的许多人迄今已成功地获得美国国立卫生研究院的赠款（7 R 01，3 COBRE，2 R21，2博士后奖学金）。此外，另外4个R 01提案最近已经过审查，并获得了有利的支持率（13%、15%、16.2%和22%），因此它们很有可能获得资助。所有这些新资助的项目都需要广泛使用共焦显微镜。显微镜系统将以几种不同的方式使用。D博士戈扎尔河刘和程将 使用共聚焦显微镜分析慢性间歇性缺氧介导的和衰老诱导的脑神经元细胞死亡、活性氧（ROS）产生、蛋白质聚集、心脏中心脏轴突和终末的重塑以及受体表达/共定位，用不同的染料标记或免疫染色。爱泼斯坦博士和Y. Liu将研究β细胞和心肌中糖尿病相关蛋白和ROS的产生。Dr. E. Gozal研究了通过缺氧调节Akt激活的蛋白质-蛋白质相互作用，并鉴定了凋亡的标志物，如响应间歇性缺氧的凋亡细胞外膜上的膜联蛋白V。王博士将对细胞内Ca 2+分布进行成像，测量线粒体膜电位，并研究缺氧诱导的亚细胞结构中凋亡蛋白的易位。Lassiter博士将研究补体膜的存在 攻击复合物（MAC）在体内沉积到缺血后的脑细胞，并确定MAC沉积后的命运。KCHRI对该显微镜系统的采购和维护有着坚定的承诺。为了支持这一应用，KCHRI将翻新一个100平方英尺的房间来容纳该系统，并将承担该仪器的长期服务合同，如果购买的话。

This proposal requests to purchase and operate a Bio-Rad Confocal Microscope in conjunction with a Nikon rE 2000U inverted microscope to enhance the rapid developing research programs in Kosair Children Hospital Research Institute (KCHRI). Since the establishment of KCHRI in 2000, this group has recruited 8 faculty members and a large number of research scientists, postdoctoral research fellows, and students with a total of 40 researchers. Many of them have thus far successfully acquired NIH grants (7 R01,3 COBRE, 2 R21,2 postdoctoral fellowships). In addition, an additional 4 R01 proposals have been recently reviewed and received favorable percentiles (13%, 15%, 16.2%, and 22%), hence they are very likely to be funded. All these newly funded projects require an extensive use of the confocal microscope. The microscope system will be employed in several different ways. Drs. D. Gozal, R. Liu, and Cheng, will use the confocal microscope to analyze chronic intermittent hypoxia-mediated and aging-induced brain neuronal cell death, reactive oxygen species (ROS) production, protein aggregation, remodeling of cardiac axons and terminals in the heart, and receptor expressions/colocalization, labeled with different dyes or immunostained. Drs Epstein and Y. Liu will examine diabetes-related proteins and ROS production in beta cells and myocardium. Dr. E. Gozal examine protein-protein interactions that regulate Akt activation by hypoxia and to identify markers of apoptosis such as Annexin V on the outer membrane of apoptotic cells in response to intermittent hypoxia. Dr. Wang will image intracellular Ca2+ distribution, measure mitochondrial membrane potentials, and study the translocation of apoptotic proteins among subcellular structures induced by hypoxia. Dr. Lassiter will study the presence of complement membrane attack complexes (MAC) deposited in vivo onto post-ischemic cerebral cells and determine the fate of MAC following deposition. The KCHRI has a strong commitment to the acquisition and maintenance of this microscope system. In support of this application, KCHRI will renovate a 100 sq ft room to house the system and will underwrite the long-term service contract for the instrument, if it is purchased.