Multiphoton Laser Scanning Microscope: Zeiss LSM710 NLO

描述（由申请人提供）：我们申请资金购买多光子激光扫描显微镜（蔡司LSM 710 NLO），作为我们自1999年以来一直使用的过时BioRad MRC-1024 MP的替代品。使用该工具的研究是我们获得两项PPG赠款（NIDDK和NHLBI）、一项NIDDK GI核心中心资助和几项RO 1的关键，所有这些都在进行中。所有BioRad多光子仪器的服务已在过去3年中停止，显微镜、扫描仪、探测器和我们用于多光子激发的旧一代Tsunami激光器（光谱物理学）的部件也已停止。更换该仪器是确保这些NIH奖项的成功实验连续性的必要步骤。这台显微镜在三个广泛的研究领域有4个主要用户和3个次要用户：心脏病，癌症和心脏病。所有的主要和次要用户都资助了NIH项目，这些项目的目标需要使用多光子显微镜来实现特定的目标。与当前仪器一样，申报的多光子系统将成为Ross研究大楼荧光显微镜核心的一部分，该中心是NIH/NIDDK GI核心中心的一部分，Donowitz博士是PI，Kovbasnjuk博士是成像核心联合主任，两人都是主要用户。这个罗斯显微镜核心(://www.hopkinsmedicine.org/confocal/）为霍普金斯科学家提供仪器和培训已有15年，并已被JHUSOM的100多个实验室使用，包括先进的成像应用，包括福斯特共振能量转移（FRET）、光漂白后荧光恢复（FRAP）、去卷积、二次谐波产生（SHG）、pH敏感性SNARF-4FD染料、Ca指示剂染料、NAD（P）H、GFP的定量成像方案以及深部组织免疫荧光方案，用户收费部分沿着NIH资金支持多光子服务合同以及我们新的蔡司LSM 510/Meta共聚焦系统和一个核心经理谁监督行政和科学的核心活动。机构支持在国家卫生研究院资金失效或重大设备故障的情况下提供备份。拟议的仪器将是霍普金斯的第二个系统，可供一般JHUSOM研究界使用。

DESCRIPTION (provided by applicant): We request funds to purchase a multiphoton laser scanning microscope (Zeiss LSM 710 NLO) as a replacement for our outdated BioRad MRC-1024 MP which we have had since 1999. Studies using this instrument were a key to us obtaining two PPG grants (NIDDK and NHLBI), a NIDDK GI Core center grant, and several RO1s, all of which are ongoing. Service of all BioRad multiphoton instruments have been discontinued for the last 3 years and parts for the microscope, scanner, detectors and our old generation of Tsunami laser for multiphoton excitation (Spectral Physics) have been discontinued as well. Replacement of this instrument is a necessary step to assure the successful experimental continuity of these NIH awards. This microscope has 4 major and 3 minor users in three broad research areas: diarrheal diseases, cancer and cardiology. All of the major and minor users have funded NIH projects, the aims of which require use of a multiphoton microscope to accomplish the specific aims. As in the case of the current instrument, the proposed multiphoton system will be a part of the Ross Research Building Fluorescent Microscopy Core which is part of an NIH/NIDDK GI Core Center, of which Dr. Donowitz is the PI and Dr. Kovbasnjuk is the Imaging Core Co-Director and both of whom are major users. This Ross Microscopy Core (://www.hopkinsmedicine.org/confocal/) has provided instruments and training for Hopkins scientists for 15 years and has been used by over 100 laboratories at JHUSOM, including advanced imaging applications which include Forster Resonance Energy Transfer (FRET), Fluorescence Recovery After Photobleaching (FRAP), deconvolution, Second Harmonic Generation (SHG), quantitative imaging protocols for pH-sensitive SNARF-4Fdye, Ca indicator dyes, NAD(P)H, GFP, as well as deep tissue immunofluorescence protocols, etc. User charges in part along with NIH funding support the service contracts for the multiphoton as well as our new Zeiss LSM 510/META confocal system and for a Core Manager who oversees both administrative and scientific Core activities. Institutional support provides back up in case of NIH funding lapses or major equipment malfunction. The proposed instrument will be only the second system at Hopkins available for the general JHUSOM research community.