Nikon C1Si Spectral Confocal Microscope

描述(由申请人提供)：申请资金用于购买现代激光扫描共聚焦显微镜系统，安装在罗德岛医院(该州最大的布朗大学附属教学医院和创伤中心)核心研究实验室部门。尼康C1硅光谱成像共聚焦显微镜将极大地促进七名NIH资助的研究人员以及布朗大学校园研究人员的研究和教学工作，布朗大学医学院的其他五家附属医院的研究人员共同组成了布朗大学医学院。在将得到该文书支持的努力中，同样值得注意的还有当地科学界新成立的四个生物医学研究卓越中心(COBRE)。我们目前有一台破旧的尼康PCM 2000共焦显微镜，它已经很好地为我们服务了八年。然而，这是一个过时的系统(在硬件和软件方面)，不再正常运行。我们的激光强度降到了大约一半的功率，并且有一些光学/图像故障是维修工程师无法纠正的。此外，总体设计和过时的软件无法满足我们所服务的研究界的需求，特别是与整体荧光灵敏度和活细胞成像相关的需求。所要求的尼康C1硅光谱成像共聚焦显微镜是一种新颖的设计，仅在去年才上市。它是一种模块化共聚焦系统，由一台TE200-E2电动显微镜组成，配备了以下组件：1)独立的扫描头；3)光谱探测器模块，允许使用32 PMT线性阵列同时采集32个独立通道的光谱数据。此外，32PMT线阵允许在2秒内采集512个光谱图像，这是该仪器独有的能力。通过在400 nm到750 nm的范围内选择10 nm、5 nm和2.5 nm的通道宽度，可以对光谱分辨率进行调制：4)激光模块，通过确保在数字化仪发送数据和重置时不存在停机时间来提高灵敏度；5)分开控制模块和监视器，从而将一个组件之间的电子干扰或振动降至最低。采集方式包括共聚焦荧光、共聚焦光谱成像、反射成像、干涉反射对比度、明场扫描透射光、相位对比度和DIC。C1Si适用于各种实验分析，包括：多种颜色实验；环境敏感探针的光谱，如pH敏感的GFP；单程FRET；通过原位染料光谱分析去除不需要的荧光。所有这些都可以在电动Z堆栈、延时模式下完成。对这笔拨款的更正范围如此之广，以至于划界是不切实际的。相关性：所要求的仪器不仅将支持本提案中描述的七个主要用户组(MUG)项目的过渡性和基础科学研究，还将支持几个生物医学研究卓越中心(COBRE)的研究人员，包括罗德岛医院癌症研究发展中心(PI是我们MUG的成员)以及布朗大学癌症信号网络COBRE、妇女和婴儿医院围产期生物学COBRE和罗杰·威廉姆斯医学中心新干细胞生物学COBRE的研究人员。该仪器还可能使布朗大学和布朗大学附属的六家教学医院的研究人员受益，这些医院参与了生物医学研究，但不是上述Cobre计划的一部分。最后，根据需要，将向我们地区其他学院/医院和大学的调查人员提供访问权限。

DESCRIPTION (provided by applicant): Funds are requested for the acquisition of a modern Laser Scanning Confocal Microscope system to be installed within the Division of Core Research Laboratories at Rhode Island Hospital (the State's largest Brown University-affiliated teaching hospital and trauma center). The Nikon C1 Si Spectral Imaging Confocal Microscope will significantly benefit the research and teaching endeavors of the seven NIH-funded investigators whose projects are detailed in this submission as well as that of Brown University campus-based investigator, and the work of researchers at the five other Brown-affiliated hospitals which collectively comprise the Brown University Medical School. Also noteworthy among the efforts that will be supported by this instrument are four newly-established Centers of Biomedical Research Excellence (COBRE) within the local scientific community. We currently have a decrepit Nikon PCM 2000 Confocal Microscope, which has served us well for eight years. However, this is an outmoded system (in terms of hardware and software) that no longer functions properly. Our laser strength is down to approximately half-power and there are optical/image faults that the service engineers have not been able to correct. Moreover, the overall design and out-of-date software make it impossible to meet the needs of the research community we serve, especially as related to overall fluorescence sensitivity and with regard to live cell imaging. The requested Nikon C1 Si Spectral Imaging Confocal Microscope is a novel design that has only become available within the last year. It is a modular confocal system consisting of a TE200-E2 Motorized Microscope quipped with the following components: 1) a separate scan head; 3) a spectral detector module that allows simultaneous spectral data acquisition across 32 separate channels using a 32 PMT linear array . Moreover, the 32 PMT linear array allows 512 spectral image acquisition in 2 seconds, a capability that is unique to this instrument. The spectral resolution can be modulated by choosing channel widths of 10nm, 5 nm, and 2.5 nm over a range from 400nm to 750 nm: 4) a laser module which increases sensitivity by insuring that there is no down time while digitizers send data and reset; 5) separate control module, and monitor, thereby minimizing electrical interference or vibration from one component to another. Acquisition modes include confocal fluorescence, confocal spectral imaging, reflection imaging, interference reflection contrast, and scanned transmitted light in brightfield, phase contrast, and DIC. The C1Si is suitable for a wide variety of experimental analyses including: multiple color experiments; spectroscopy of environmentally sensitive probes like pH sensitive GFP; single pass FRET; removal of unwanted fluorescence by spectral analysis of dyes in-situ. All of the above can be done in motorized Z stack, time lapse mode. Corrections to this grant are so extensive that demarcations are impractical. Relevance: The requested instrumentation will not only support the transitional and basic science research of the seven Major User Group (MUG) projects described in this proposal, it will also support investigators of several Centers Of Biomedical Research Excellence (COBRE) including Rhode Island Hospital's COBRE For Cancer Research Development (the PI of which is a member of our MUG) as well as investigators of Brown University's COBRE For Cancer Signaling Networks, the Women and Infants' Hospital COBRE For Perinatal Biology, and the Roger Williams Medical Center COBRE for New Stem Cell Biology. The instrumentation will also potentially benefit investigators from Brown University and the six Brown University-affiliated teaching hospitals that are involved in biomedical research but are not part of the above COBRE programs. Finally, access will be provided to investigators from other colleges/hospitals, and universities in our area as needed.