Multiphoton Laser Scanning Microscope: Zeiss LSM710 NLO

描述(由申请人提供)：我们请求资金购买多光子激光扫描显微镜(蔡司LSM 710NLO)，以取代我们自1999年以来一直拥有的过时的BioRad MRC-1024 MP。使用该仪器的研究是我们获得两项PPG资助(NIDDK和NHLBI)、NIDDK GI核心中心资助和几个RO1的关键，所有这些都在进行中。所有BioRad多光子仪器的服务在过去3年都已停止，显微镜、扫描仪、探测器和我们用于多光子激发(光谱物理)的老一代海啸激光器的部件也已停止使用。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：腹泻疾病、癌症和心脏病学。所有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目标。与目前的仪器一样，拟议的多光子系统将是罗斯研究大楼荧光显微镜核心的一部分，罗斯研究大楼荧光显微镜核心是NIH/NIDDK GI核心中心的一部分，多诺维茨博士是该中心的PI，Kovbasnjuk博士是成像核心的联席主任，两人都是主要用户。罗斯显微镜核心(www.HopkinsMedicine.org/conocus/)已经为霍普金斯大学的科学家提供了15年的仪器和培训，并已被JHUSOM的100多个实验室使用，包括先进的成像应用，包括Forster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snarf-4F染料的定量成像协议、钙指示剂染料、NAD(P)H、GFP以及深层组织免疫荧光协议。用户收费部分与NIH的资金一起支持多光子和我们新的蔡司LSM 510/Meta共焦系统的服务合同，以及监督行政和科学核心活动的核心经理的服务合同。在NIH资金短缺或重大设备故障的情况下，机构支持提供后备。拟议的仪器将是霍普金斯大学为普通JHUSOM研究界提供的第二个系统。

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We request funds to purchase a multiphoton laser scanning microscope (Zeiss LSM 710 NLO) as a replacement for our outdated BioRad MRC-1024 MP which we have had since 1999.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.