Multiphoton laser scanning microscope

多光子激光扫描显微镜

• 批准号: 6580480
• 负责人: Ben W Strowbridge
• 金额: $ 49.93万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6580480
• 关键词: biomedical equipment purchase; brain imaging /visualization /scanning; lasers; microscopy

DESCRIPTION (provided by applicant): Funds are requested to purchase integrated multiphoton laser scanning microscope system for imaging living brain tissue. This non-linear optical (NLO) system will be based on the Zeiss LSM 510 scan system and an Axioskop 2Plus FS motorized upright microscope. The primary fluorescent excitation source will be a 5 watt Coherent titanium-sapphire femtosecond laser. This system will be optimized for simultaneous NLO imaging and patch clamp electrophysiology in living brain slices, nematodes and semi-intact spinal cord preparations. The proposed instrument will be housed and administered by the Neuroscience Imaging Center (NIC). The NIC currently maintains two other major microscope systems and has a full-time technician available for user training and minor maintenance. Projects from ten users (four major and six minor) are described. All proposed projects exploit the unique advantages of this upright NLO system including reduced phototoxicity, deep optical sectioning through thick living preparations, and simultaneous whole cell patch clamp electrophysiology. Dr. Ben Strowbridge will use NLO imaging to record unitary presynaptic calcium transients in olfactory bulb and dentate gyrus brain slices. Dr. Lynn Landmesser will use NLO imaging to study local circuits in the developing spinal cord in chick embryos and transgenetic mice. Dr. Stefan Herlitze plans to use this multiphoton microscope to study calcium channel function in living C. elegans. Dr. David Friel will use NLO imaging to study subcellular calcium handling in Purkinje neurons in cerebellar brain slices from normal and mutant mice.

描述（申请人提供）：申请资金购买集成多光子激光扫描显微镜系统，用于成像活体脑组织。该非线性光学（NLO）系统将基于Zeiss LSM 510扫描系统和Axioskop 2 Plus FS电动立式显微镜。主要荧光激发源为5瓦相干钛-蓝宝石飞秒激光器。该系统将针对活体脑切片、线虫和半完整脊髓制备物中的同时NLO成像和膜片钳电生理学进行优化。申报仪器将由神经科学成像中心（NIC）存放和管理。NIC目前维护另外两个主要的显微镜系统，并有一名全职技术人员负责用户培训和小型维护。从十个用户（四个主要和六个次要）的项目进行了说明。所有拟议的项目都利用了这种直立式NLO系统的独特优势，包括降低光毒性，通过厚的活体制备进行深层光学切片，以及同时进行全细胞膜片钳电生理学。Ben Strowbridge博士将使用NLO成像记录嗅球和齿状回脑切片中的单一突触前钙瞬变。林恩兰德梅瑟博士将使用NLO成像研究鸡胚和转基因小鼠脊髓发育中的局部回路。Stefan Herlitze博士计划使用这种多光子显微镜来研究活体C细胞中钙通道的功能。优雅的大卫弗里尔博士将使用非线性光学成像研究亚细胞钙处理浦肯野神经元在小脑脑切片从正常和突变小鼠。