Acquisition of Zeiss 710 Confocal Laser Scanning Microscope

描述（由申请人提供）：乔治华盛顿大学医学中心（GWUMC）的显微镜和图像分析中心（CMIA）为获取和处理与生物医学科学相关的显微镜图像提供核心资源。1999年，在承诺提供大量机构支持的情况下，CMIA成为机构核心基金。它一直忠于其最初的目标，研究，传播知识，并在几个层面提供教育机会。CMIA提供过时的BioRad MRC 1024 ES共聚焦激光扫描显微镜，由于缺乏服务合同，其维护受到严重挑战。我们现在寻求获得下一代共聚焦激光扫描显微镜(CLSM) -蔡司LSM 710。所要求的CLSM将配备多种激光激发协议和光谱发射检测，以及环境控制，以解决当前设备的缺陷，并满足NIH资助研究人员的研究目标。与现有的CLSM相比，该仪器除了具有更高的检测效率和可靠性外，还将能够进行激光刺激，从而实现复杂的可视化方法，如荧光共振能量转移（FRET）、光漂白和光转换。光谱检测将为研究人员在选择发射带宽、消除串扰和自身荧光方面提供更高水平的灵活性。额外的激光线（405nm）将为最先进的标记（如量子点）提供标记检测。拟议的仪器将配备环境控制，结合激发和发射协议提供的最大灵活性，将推进研究人员的生命细胞成像实验-这是目前仪器无法做到的。CLSM将是该机构同类仪器中的第一台，通过使用线性非混合算法，即使有重叠的发射光谱，也能够同时检测多达八个荧光团的光谱特征。提出的CLSM将促进荧光标记检测，将为我们提供研究蛋白质-蛋白质相互作用的能力，并将使我们能够应用新的和更可靠的标记策略。一组建立良好的NIH资助的调查人员支持这一申请。这些研究人员将能够超越现有BioRad 1024ES CLSM所施加的限制来推进他们的项目。CMIA将利用其多用户环境来宣传研究人员与拟议的CLSM的成就，这将为所有NIH资助的研究人员创建一个广泛的方法共享和进步平台。因此，该文书将促进部门间的相互作用和合作。我们的生物医学科学研究所博士项目的学生可以在研究轮转和论文项目中使用该仪器进行培训。公共卫生相关性：乔治华盛顿大学医学中心核心设施中可用的共聚焦激光扫描显微镜所造成的缺陷在本应用中得到解决。建议购买最先进的共聚焦激光扫描显微镜，这将集成到核心设施中。新仪器将允许一组成熟的研究人员推进他们的NIH资助的项目。

DESCRIPTION (provided by applicant): The Center for Microscopy and Image Analysis (CMIA) at The George Washington University - Medical Center (GWUMC) provides core resources for acquisition and processing of microscopic images of relevance to biomedical sciences. The CMIA was made an institutional core facility in 1999 by a commitment of considerable institutional support. It has remained true to its original goals of research, dissemination of knowledge, and educational opportunities at several levels. CMIA offers outdated BioRad MRC 1024 ES confocal laser scanning microscopes, the maintenance of which is seriously challenged by the lack of service contracts. We now seek to acquire the next generation of confocal laser scanning microscope (CLSM) - Zeiss LSM 710. The requested CLSM will be equipped with variety laser excitation protocols and spectral emission detection, and environmental control to address current equipment deficiencies and to meet the research objectives of NIH funded investigators. In addition to the significantly higher detection efficiency and reliability compared to the existing CLSM, the proposed instrument will be capable of laser stimulation, allowing the implementation of sophisticated visualization methods such as fluorescent resonance energy transfer (FRET), photobleaching and photoconversion. Spectral detection will provide the researchers with a higher level of flexibility in selecting the emission bandwidth, elimination of cross-talk and autofluorescence. Additional laser lines (405nm) will offer labeling detection for state of the art markers such as quantum dots. Proposed instrument will be equipped with environmental control, which in combination of the greatest flexibility offered by excitation and emission protocols will advance investigators life cell imaging experiments - not possible with the current instrument. The CLSM will be the first of its class in the institution and will be capable of simultaneously detecting spectral signatures from up to eight fluorophores even if they have overlapping emission spectra, by using linear un-mixing algorithms. Proposed CLSM will advance fluorescence labeling detection, will provide us with the capability for studying protein-protein interactions, and will allow us for applying new and more reliable labeling strategies. A group of well established NIH funded investigators support this application. These investigators will be able to advance their projects beyond the limits imposed by the existing BioRad 1024ES CLSM. CMIA will use its multi-user environment for publicizing the achievements of investigators with the proposed CLSM, which will create a broad platform for methodological sharing and advancement of all NIH funded investigators. Thus, the instrument will foster interdepartmental interactions and collaborations. Training with the instrument will be available to our Institute of Biomedical Sciences PhD program students on research rotations and dissertation projects. PUBLIC HEALTH RELEVANCE: The deficits imposed by the available confocal laser scanning microscope available at The George Washington University Medical Center core facility are addressed in this application. Acquisition of a state of the art confocal laser scanning microscope is proposed, which will be integrated in into the core facility. The new instrument will allow a group of well established investigators to advance their NIH funded project.