LSM CONFOCAL SYSTEM: DEVELOPMENTAL BIOLOGY

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，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，以购买蔡司LSM 510 Meta NLO的直立落射荧光Axioplan 2显微镜。在过去的几年里，多光子（双光子激光扫描）显微镜的发明已经引领细胞研究进入新的领域，提供活动物体内细胞形态，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，因此能够对组织内的深层（数百微米）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（数小时至数周）重复成像。这是因为激发被有效地限制在光束的焦点，使得上方和下方的荧光团不被激发。光漂白和光毒性，共聚焦显微镜的共同局限性，从而大大减少使用多光子显微镜时。该显微镜将用于支持华盛顿大学两个不同系的三名主要研究人员正在进行的资助研究计划。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疾病，眼科和视觉研究以及耳聋和听觉功能障碍的研究产生影响。所要求的仪器将被安置在生物结构成像设施，并将由该部门的资金管理和维护。拟议的行政结构和仪器的管理进行了优化，以最好地利用显微镜的时间，从而确保最大的利益，以主要研究人员，以及广泛的二级用户在大学。

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.