Multi-photon imaging system

描述（由申请人提供）：多光子显微镜可能是自共聚焦显微镜引入以来荧光显微镜最重要的进步。多光子显微镜允许可视化深入到结构，并允许这种可视化在延长的时间内进行，对标本的损害最小。由多光子显微镜提供的优点包括：（a）没有荧光团的焦点外漂白;（B）更深的光学成像，因为减少了焦点外荧光团激发和减少了入射波长的散射;以及（c）减少了活样本的光毒性。此应用程序要求资金购买：1.）BioRad Radiance 2100多光子系统; 2.）相干Mira 900-F可调谐Ti：S激光器和Verdi 8瓦ND：YVO 4激光器;以及3.）尼康E-800正置显微镜和尼康TE-2000 U倒置显微镜。选择Radiance 2100多光子系统的原因有很多，其中之一是扫描头可以相对轻松地从一个显微镜支架移动到另一个显微镜支架，允许成像系统与直立支架一起用于体内成像或与倒置支架一起用于体外成像。这是本应用中的一个重要考虑因素，因为多光子系统将安装在校园范围的显微镜设施中，并为具有不同研究兴趣和需求的广泛用户提供服务。Radiance 2100多光子成像系统将安装在W.M. Kcck生物成像实验室该实验室成立于1993年，由W。M. Keck基金会，并配备有BioRad MRC 1024共聚焦显微镜;具有混合气体、氪/氩激光器的单光子显微镜，能够三线激发。凯克实验室积极管理，由用户费用支持，是威斯康星大学麦迪逊分校校园内唯一的共聚焦显微镜服务资源。多光子成像系统将由凯克实验室操作和维护，成像系统将向所有合格用户开放，但须遵循自实验室成立以来成功运作的既定程序。目前，该实验室提供了50多名教师和他们的学生在生物学的不同领域的机会，使用共聚焦显微镜在他们的研究。在这些教师和学生所代表的广泛的研究领域是：农学、动物科学、生物医学工程、细胞、发育和分子生物学、遗传学、神经科学、营养科学、肿瘤学、病理学、生殖、心脏和肌肉生理学等。如果凯克实验室也提供多光子成像系统，那么这些用户的研究计划将得到显著的推进。如上所述，BioRad 2100多光子成像系统和相关设备的收购将补充Kcck实验室的BioRad MRC 1024单光子共聚焦显微镜，并将为来自整个UW-Madison校园的生物科学家开辟新的研究途径。

DESCRIPTION (provided by applicant): Multi-photon microscopy is perhaps the most important advance in fluorescence microscopy since the introduction of the confocal microscope. Multi-photon microscopy allows visualization deep into structures and permits this visualization to be carded out over extended periods of time with minimal damage to the specimen. Among the advantages provided by multi-photon microscopy are: (a) no out of focus bleaching of fluorophores; (b) deeper optical imaging, because of reduced out of focus fluorophore excitation and decreased scattering of incident wavelengths; and (c) reduced phototoxicity for living specimens. This application requests funds to purchase a: 1.) BioRad Radiance 2100 multi-photon System; 2.) Coherent Mira 900-F tuneable Ti:S laser and Verdi 8 watt ND:YVO4 laser; and 3.) Nikon E-800 upright microscope and a Nikon TE-2000U inverted microscope. The Radiance 2100 multi-photon system has been selected for many reason, among them being a scanhead that can be moved with relative ease from one microscope stand to another, allowing the imaging system to be used with an upright stand for in vivo imaging or with an inverted stand for in vitro imaging. This is an important consideration in this application, because the multi-photon system will be installed in a campus-wide microscopy facility and serve a wide-range of users with different research interests and needs. The Radiance 2100 multi-photon imaging system will be housed in the W.M. Kcck Laboratory for Biological Imaging. The Laboratory was established in 1993 with a gift from the W. M. Keck Foundation, and is equipped with a BioRad MRC 1024 confocal microscope; a single-photon microscope with a mixed gas, krypton/argon laser, capable of three-line excitation. The Keck Laboratory is actively managed, supported by user fees, and is the only confocal microscopy service resource on the UW-Madison campus. The multi-photon imaging system will be operated and maintained by the Keck Laboratory, and access to the imaging system will be open to all qualified users, following well-established procedures that have functioned successfully since the inception of the Laboratory. Currently, the Laboratory provides more than 50 faculty and their students in different areas of biology the opportunity to use confocal microscopy in their research. Among the broad research areas represented by these faculty and students are: agronomy, animal science, biomedical engineering, cell, developmental, and molecular biology, genetics, neuroscience, nutritional sciences, oncology, pathology, reproductive, cardiac, and muscle physiology, etc. The research programs of many of these users will be advanced significantly if a multi-photon imaging system also is available to them in the Keck Laboratory. The acquisition of the BioRad 2100 multi-photon imaging System and associated equipment, as described above, will complement the Kcck Laboratory's BioRad MRC 1024 single-photon confocal microscope, and it will open new avenues of investigation for biological scientists from across the UW-Madison campus.