Two Photon Microscope

描述（由申请人提供）：本申请要求Zeiss LSM 510 NLO双光子显微镜，该显微镜针对活细胞和组织的定量成像进行了优化。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，连接到具有盖玻片校正水浸物镜的Zeiss Axiovert 200显微镜。该设备将被纳入一个新的活显微镜核心设施，将服务于来自整个辛辛那提大学医学院多个部门的研究人员，他们需要该仪器来执行超出共聚焦显微镜能力的活细胞和活组织荧光显微镜。我们的4个主要用户群需要多光子显微镜（1）在距表面大于50 mu/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），和（2）允许能量的局部沉积，用于解开，光损伤，或者光漂白实验，这样就可以在同一个显微镜上用传统的共聚焦显微镜来跟踪事件。所有主要用户都证明，基于蔡司平台的多光子显微镜将在满足他们的实验需求方面提供超越共焦显微镜的重大进步。我们的用户的需求不能满足我们机构的设备，因为在整个大学没有多光子显微镜，共聚焦显微镜是不足以解决用户的问题。多光子显微镜将安装在PI实验室附近的位置，这是所有研究者的中心，并将有助于PI对设备状态进行日常监测。该申请得到了大学的大量兴趣，需求和财政支持的支持，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.