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

描述(由申请人提供)：马里兰大学巴尔的摩分校(UMB)的首席研究人员迫切需要奥林巴斯Viva-View孵化器显微镜系统来监控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六名研究人员的十二项NIH R01拨款，以及其他拨款。由于显微镜是在培养箱内设计的，并包括更换培养基口，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。该显微镜使用DIC或表观荧光光学元件来获取图像、电影、Z-堆栈等。此功能非常适合监控细胞形状、细胞内结构和融合蛋白的细胞内分布的变化，以及荧光指示剂在细胞内的分布(例如，Indo-1双发射钙离子成像)，时间从几小时到几周不等。为了促进科学交流、促进研究合作和增加用户可用性，孵化器中的显微镜系统将被整合到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位于校园的一个偏远角落，而BMBIC的开发是为了满足校园这一象限的调查仪器需求。最近对BMBIC空间的翻新表明了马里兰大学医学院和生物化学和分子生物学系对该设施的承诺。所有主要用户的一个关键的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。新的孵化器显微镜将使这一潜力成为可能。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是重要的。事实上，提供使用最先进的仪器设备是该局培训任务的一个关键方面。新的孵化器显微镜将加强至少两项由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控培养的成人骨骼肌纤维的去分化、骨骼肌管和肌纤维的发育和形成、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孵化器来支持该仪器。对BMBIC的一般监督由BMBIC监督委员会提供，并为该文书指派了一个特定的监督小组。仪器维护由医学院、生物化学和分子生物学系以及使用费提供支持。

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.