HIGH-SPEED CONFOCAL MICROSCOPE FOR A SHARED FACILITY

适用于共享设施的高速共焦显微镜

• 批准号: 2489110
• 负责人: William Jonathan Lederer
• 金额: $ 39.02万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-18 至 2000-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2489110
• 关键词: biomedical equipment; biomedical equipment purchase; confocal scanning microscopy; electrophysiology; photolysis

Cells control many of their functions through localized (micron to sub- micron scale) concentration changes of intracellular messengers. These changes often occur rapidly (millisecond to sub-millisecond scale). For example, contraction of muscle cells and section from neurons are regulated by transient (1-100 ms) increases in intracellular calcium. The introduction of calcium-sensitive fluorescent indicators like fluo-3 discovered in part by J. Kao (Project I) while working in the laboratory of R.Y. Tsien, opened the way for microscopical studies of calcium transients in living cells. Coupled with the improved spatial resolution of confocal microscopy, calcium imaging studies made it possible to observe the release of calcium from single channels or single "release units" in intracellular stores. These 'calcium sparks' were first discovered by Lederer (Project II) in cardiac (Cheng et al., 1993) and smooth muscle (Nelson et al., 1995) cells, and were subsequently discovered in skeletal muscle cells in collaboration with other campus researchers (Klein et al., 1996). These and related studies have contributed to the University of Maryland's stature as a major research center for studies of intracellular calcium signaling and regulation, which is the primary research focus of more than 20 independent campus laboratories. Further advances in this area require improvements in temporal resolution so that the kinetics of spark formation and propagation, and their relationship to the calcium transients they evoke, can be characterized. The equipment requested by this application is intended to create a multi- user core facility that meets this requirement. A high-speed confocal microscope and dedicated voltage clamp apparatus will enable researchers at the University of Maryland and other area institutions to initial and examine calcium sparks with high spatial and temporal resolution. An integrated photolysis laser and temperature-controlled perfusion system will enable them to manipulate the intracellular and extracellular environment of living cells to allow correlation betweens parks and various stimuli. In addition to enabling new approaches to the study of calcium sparks and other dynamic phenomena in living cells, this core facility will have a significant impact on the teaching mission of the University of Maryland.

细胞通过局部（微米到亚微米）控制它们的许多功能。 微米尺度）细胞内信使的浓度变化。这些 变化经常迅速发生（毫秒到亚毫秒尺度）。为 例如，肌肉细胞的收缩和神经元的切片， 由细胞内钙的瞬时（1-100 ms）增加调节。的 引入钙敏感荧光指示剂，如Fluo-3 在实验室工作时，J. Kao（项目I）发现了部分 关于R.Y.钱存训开创了钙的显微镜研究 活细胞中的瞬变现象。再加上空间分辨率的提高 共聚焦显微镜，钙成像研究使得有可能 观察钙从单个通道或单个“释放”的释放 单位”的细胞内商店。这些“钙火花”首先是 由Lederer（Project II）在心脏中发现（Cheng等人，（1993年）和 平滑肌（纳尔逊等人，1995）细胞，随后 在骨骼肌细胞中发现， 研究人员（Klein等人，1996年）。这些研究和相关研究 有助于马里兰州的地位，作为一个主要的研究 细胞内钙信号和调节研究中心， 这是20多个独立校区的主要研究重点 laboratories. 这一领域的进一步发展需要改进时间分辨率 因此，火花形成和传播的动力学及其 与它们引起的钙瞬变的关系，可以表征。 本申请所要求的设备旨在创建一个多- 用户核心设施，满足这一要求。一种高速共焦 显微镜和专用电压钳装置将使研究人员 在马里兰州大学和其他地区的机构， 以高空间和时间分辨率检查钙火花。一个 集成光解激光和温控灌注系统 将使他们能够操纵细胞内和细胞外 活细胞的环境，使公园和 各种刺激。除了能够采用新的方法来研究 钙火花和活细胞中的其他动态现象，这个核心 该设施将对教学使命产生重大影响， 马里兰州大学。