Multi-photon imaging system

描述(申请人提供)：自共焦显微镜问世以来，多光子显微镜可能是荧光显微镜中最重要的进步。多光子显微镜允许深入结构的可视化，并允许这种可视化在较长的一段时间内进行，对标本的损害最小。多光子显微镜提供的优点包括：(A)荧光团不会发生焦点外漂白；(B)由于焦点外荧光团激发和入射波长散射减少，光学成像更深入；以及(C)对活标本的光毒性降低。此应用程序请求资金购买：1。)Biorad Radiance 2100多光子系统；相干米拉900-F可调谐钛：S激光器和威尔第8瓦Nd：YVO_4激光器；尼康E-800立式显微镜和尼康TE-2000U倒置显微镜。Radiance 2100多光子系统的选择有很多原因，其中之一是扫描头可以相对容易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，或者与倒置支架一起用于体外成像。这是这一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内的显微镜设施中，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。该实验室是在W.M.Keck基金会的捐赠下于1993年建立的，配备了一台BioRad MRC1024共焦显微镜；一台具有混合气体、氪/Ar激光的单光子显微镜，能够进行三线激发。凯克实验室积极管理，由用户付费支持，是威斯康星大学麦迪逊分校唯一的共焦显微镜服务资源。多光子成像系统将由凯克实验室操作和维护，所有符合条件的用户都可以使用该成像系统，遵循自该实验室成立以来成功运作的既定程序。目前，该实验室为生物学不同领域的50多名教师和他们的学生提供了在他们的研究中使用共焦显微镜的机会。这些教师和学生代表的广泛研究领域包括：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。如果凯克实验室也向这些用户提供多光子成像系统，他们的研究计划将大大进步。如上所述，收购BioRad 2100多光子成像系统和相关设备将补充Kcck实验室的BioRad MRC1024单光子共焦显微镜，并将为威斯康星大学麦迪逊分校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.