Two Photon Microscope

描述（由申请人提供）：本申请需要一台蔡司LSM 510 NLO双光子显微镜，用于活细胞和组织的定量成像。钛蓝宝石激光器（相干变色龙扩展范围）将直接耦合到扫描头的输入端口，连接到蔡司Axiovert 200显微镜，具有盖盖校正的水浸物镜。该设备将被整合到一个新的活体显微镜核心设备中，该设备将为来自辛辛那提大学医学院多个部门的研究人员提供服务，他们需要该仪器进行活细胞和活组织荧光显微镜，这超出了共聚焦显微镜的能力。我们的4个主要用户群体需要多光子显微镜来(1)在距离表面大于50mu/m的焦深处提供组织内细胞和亚细胞事件的高分辨率成像（在这些深度，共聚焦显微镜既不能提供高空间分辨率，也不能避免光损伤），(2)允许局部沉积能量，以去除细胞，光损伤，或者光漂白实验，这样就可以在同一台显微镜上用传统的共聚焦显微镜追踪事件。所有主要用户都记录了基于蔡司平台的多光子显微镜将在解决他们的实验需求方面提供超越共聚焦显微镜的重大进步。我们的设备不能满足我们的用户的需求，因为整个大学没有多光子显微镜，共聚焦显微镜不足以解决用户的问题。多光子显微镜将安装在PI实验室附近的一个地点，这是所有调查人员的中心，并将便利PI每天监测设备状态。该申请得到了大学的大量兴趣、需求和财政支持，并将为nih资助的科学界提供强有力的服务。

DESCRIPTION (provided by applicant): This application requests a Zeiss LSM 510 NLO two-photon microscope optimized for quantitative imaging with living cells and tissues. A Titanium-sapphire laser (Coherent Chameleon extended range) will be directly coupled to the input port of the scan head, attached to a Zeiss Axiovert 200 microscope having coverslip-corrected water immersion objectives. The device will be incorporated in a new Live Microscopy Core facility that will serve investigators from multiple departments throughout the University of Cincinnati College of Medicine, who need the instrument to perform live cell and live tissue fluorescence microscopy that is beyond the capabilities of confocal microscopy. Our 4 major user groups require multi-photon microscopy to (1) provide high-resolution imaging of cellular and subcellular events within tissue at focal depths greater than 50mu/m from the surface (at these depths, confocal microscopy can neither provide high spatial resolution nor avoid photodamage), and (2) permit localized deposition of energy for uncaging, photodamage, or photobleaching experiments so that events can then be tracked by conventional confocal microscopy on the same microscope. All major users have documented that multi-photon microscopy based on the Zeiss platform will provide a significant advance beyond confocal microscopy in addressing their experimental needs. The needs of our users can not be met by equipment at our institution, because there is no multi-photon microscope at the entire University, and confocal microscopy is inadequate to address the questions of the users. The multi-photon microscope would be installed in a location near the laboratory of the PI, which is central to all investigators, and would facilitate daily monitoring of equipment status by the PI. The application is supported by substantial interest, need, and financial support at the University, and will serve a strongly NIH-funded scientific community.