In-Incubator Live Cell Fluorescence/DIC Time Lapse Imaging System

描述（由申请人提供）：巴尔的摩马里兰大学 (UMB) 的主要研究人员迫切需要奥林巴斯 Viva-View 培养箱内显微镜系统来监测细胞和组织功能和形态的长期变化。该系统将支持授予马里兰大学 SOM 六名研究人员的 12 项 NIH R01 资助以及其他资助。由于显微镜设计在培养箱内，并包含用于更换培养基的端口，因此可以在恒定的温度和气体条件下长时间观察细胞培养物。该显微镜使用 DIC 或落射荧光光学器件来采集图像、视频、z 堆栈等。此功能非常适合在数小时到数周内监测细胞形状、细胞内结构、融合蛋白的细胞内分布以及荧光指示剂的细胞内分布（例如 indo-1 双发射 Ca2 成像）的变化。为了促进科学交流、刺激研究合作并提高用户可用性，培养箱内显微镜系统将被纳入位于生物医学研究设施 (BRF) 的现有生物化学和分子生物学仪器核心 (BMBIC) 中。 BRF 位于校园的偏远角落，BMBIC 的开发是为了支持校园这一象限的研究人员仪器需求。 BMBIC 空间最近的翻新表明了马里兰大学医学院和生物化学与分子生物学系对该设施的承诺。所有主要用户的一个关键长期研究目标是在分子水平上了解与人类疾病有关的长期生物事件。新型培养箱显微镜将发挥这一潜力。我们认为，对于 UMB 的学生和博士后来说，接触广泛的成像技术并学习如何监测细胞功能的长期变化非常重要。事实上，提供最先进的仪器是 UMB 培训任务的一个关键方面。新的培养箱内显微镜将加强对至少两项 NIH 资助的培训补助金（T32GM066706-05 和 5T32AR007592-13）支持的学生的培训。该系统将使研究能够解决与调节培养成体骨骼肌纤维去分化、骨骼肌管和肌纤维发育和形成、表面上皮细胞分层和分化、乳腺癌细胞行为、DNA损伤修复和摄取以及神经元细胞外信号分子的细胞内重新分布相关的关键问题。 BMBIC 中提供了组织培养罩和标准组织培养箱来支持该仪器。 BMBIC 监督委员会负责对 BMBIC 进行总体监督，并为此工具指定了一个特定的监督小组。仪器维护由医学院、生物化学与分子生物学系和用户费用支持。

DESCRIPTION (provided by applicant): Principal investigators at the University of Maryland, Baltimore (UMB) have a pressing need for an Olympus Viva-View In-Incubator Microscope System to monitor long-term changes in cell and tissue function and morphology. This system will support twelve NIH R01 grants awarded to six investigators at the University of Maryland SOM, as well as other grants. Because the microscope is designed inside an incubator and includes port for media change, cell cultures can be viewed over long periods under constant conditions of temperature and gas. The microscope uses DIC or epi-fluorescence optics to acquire images, movies, z- stacks, etc. This capability is ideal for monitoring changes in cell shape, intracellular structure, and intracellular distribution of fusion proteins, and intracellular distribution of fluorescent indicators (e.g., indo-1 dual emission Ca2+ imaging) at times ranging from hours to weeks. To promote scientific exchange, stimulate research collaborations, and increase user availability, the in-incubator microscope system will be incorporated into the existing Biochemistry and Molecular Biology Instrumentation Core (BMBIC) located in the Biomedical Research Facility (BRF). The BRF is located in a remote corner of campus and the BMBIC was developed to support investigator instrument needs in this quadrant of campus. Recent renovation of BMBIC space indicates the commitment of the University of Maryland School of Medicine and the Department of Biochemistry and Molecular Biology to this facility. A key long range research goal of the all major users to understand long-term biological events at the molecular level as it pertains to human disease. The new in- incubator microscope will make this potential available. We believe it is important for students and postdoctoral associates at UMB to gain exposure to a wide range of imaging technology and to learn how to monitor long-term changes in cell function. Indeed, providing access to state-of-the-art instrumentation is a key aspect of the training mission at UMB. The new in-incubator microscope will enhance the training of students supported by at least two NIH-funded training grants (T32GM066706-05 and 5T32AR007592-13). This system will enable research to address key questions relating to events regulating dedifferentiation of cultured adult skeletal muscle fibers, skeletal myotube and myofiber development and formation, surface epithelial cell stratification and differentiation, breast cancer cell behavior, DNA damage repair and uptake and intracellular redistribution of extracellular signaling molecules in neurons. A tissue culture hood and standard tissue culture incubator are available in the BMBIC to support this instrument. General oversight for the BMBIC is provided by the BMBIC Oversight Committee and a specific oversight group is assigned to this instrument. Instrument maintenance is supported by the School of Medicine, the Department of Biochemistry and Molecular Biology, and users fees.