LSM CONFOCAL SYSTEM: EYE AND VISION

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：我们正在申请资金购买蔡司LSM 510 META NLO，用于直立式epifluorescence Axioplan 2显微镜。在过去的几年里，多光子（双光子激光扫描）显微镜的发明使细胞研究进入了新的领域，提供了活体动物细胞形态、功能和发育的实时视图。多光子显微镜使用高能红外脉冲激发荧光团，因此能够对组织内深层（数百微米）的结构进行成像。多光子成像的另一个优点是，活体标本可以在很长一段时间内（数小时到数周）反复成像。这是因为激发被有效地限制在光束的焦点上，这样上面和下面的荧光团就不会被激发。因此，使用多光子显微镜时，共聚焦显微镜常见的局限性光漂白和光毒性大大减少。这台显微镜将用于支持华盛顿大学两个不同部门的三位主要研究人员正在进行的资助研究项目。虽然这些研究将侧重于神经发育和可塑性的基本机制，但这些发现将对神经科学和神经系统疾病、眼科学和视觉研究以及耳聋和听觉功能障碍的研究具有重要意义。所要求的仪器将安置在生物结构成像设施内，由该部提供的资金管理和维护。该仪器的行政结构和管理经过优化，以最大限度地利用显微镜时间，从而确保主要研究人员的最大利益，以及大学广泛的次要用户。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): We are requesting funds to purchase a Zeiss LSM 510 META NLO on an upright epifluorescence Axioplan 2 microscope. Over the last several years, the invention of multiphoton (two-photon laser scanning) microscopy has spearheaded cellular research into new realms, providing real-time views of cell morphology, function and development within the living animal Multiphoton microscopy uses high-energy infrared pulses to excite fluorophores, therefore enabling imaging of structures deep (hundreds of micrometers) within the tissue. Another advantage of multiphoton imaging is that living specimens can be imaged repeatedly over long periods of time (hours to weeks). This is because excitation is effectively confined to the focal point of the beam, such that fluorophores above and below are not excited. Photobleaching and phototoxicity, common limitations of confocal microscopy, are thus much reduced when using multiphoton microscopy. This microscope will be used to support the ongoing, funded research programs of three primary investigators in two different departments at the University of Washington. Although these studies will focus on basic mechanisms of neural development and plasticity, the findings will have implications for research in neuroscience and neurological diseases, ophthalmology and vision research and deafness and auditory dysfunction. The requested instrument would be housed in the Biological Structure Imaging facility, and would be administered and maintained by funds from the department. The proposed administrative structure and management of the instrument are optimized for the best utilization of the microscope time, thus assuring the maximum benefit to the primary investigators, as well as a broad range of secondary users at the University.