LSM Confocal System

描述(由申请人提供)：我们正在申请资金购买蔡司LSM 510 Meta NLO，安装在直立的荧光Axioplan 2显微镜上。在过去的几年里，多光子(双光子激光扫描)显微镜的发明将细胞研究带入了新的领域，提供了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来激发荧光团，从而能够对组织内深部(数百微米)的结构进行成像。多光子成像的另一个优点是，活的标本可以在很长一段时间内(几小时到几周)重复成像。这是因为激发实际上被限制在光束的焦点上，因此上面和下面的荧光团都不会被激发。因此，当使用多光子显微镜时，光漂白和光毒性，这是共聚焦显微镜的常见局限性，可以大大减少。这台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、由他们资助的研究项目。尽管这些研究将集中在神经发育和可塑性的基本机制上，但这些发现将对神经科学和神经疾病、眼科和视力研究以及耳聋和听觉功能障碍的研究产生影响。所要求的仪器将存放在生物结构成像设施中，并将由该部提供的资金进行管理和维护。为最大限度地利用显微镜时间，对拟议的仪器行政结构和管理进行了优化，从而确保了初级研究人员以及大学范围广泛的二手用户的最大利益。

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.