ImageStream X Analyzer 2011

描述(由申请者提供)：NIH NCRR资金在此重新提交购买5激光2相机阿姆尼斯ImageStreamX与三(20x，40x，60x)物镜。这个新的平台是安尼斯大约11年前推出的原始乐器的一个极大的改进、改进和更强大的版本。这台有制度支持和专业监督的仪器将被放置在首席研究员现有的病理流式细胞术核心部门，该部门已经成功运作了近30年。所要求的仪器将填补通过共聚焦显微镜获得的高细节(但获取速度要慢得多)细胞图像与通过基于免疫荧光的多参数流式细胞术获得的无图像但快速获取的数据之间的空白。ImageStreamX结合了这两种后来的技术，提供了一种协同优势，而不是串联使用的每种技术。所要求的仪器可以量化荧光细胞探针的强度和细胞内位置，并可以以高速(高达~2000个细胞/秒)进行成像，使我们的马萨诸塞州综合医院(MGH)研究人员联盟能够分析稀有的亚群和高度异质的样本，并获得可靠和客观的统计结果。这项技术将使一些原本不可能进行的调查得以实际实施。通过将流式细胞术的速度、灵敏度、免疫表型鉴定能力和能力与显微镜的详细图像和功能洞察力相结合，所需的仪器将成为波士顿市中心和查尔斯敦麻省理工学院研究园区以及整个大伙伴医疗保健系统的代表研究人员的宝贵补充。该仪器使用荧光显微镜和流式细胞术中使用的相同染料和标记，几乎可以执行任何标准的流式细胞仪分析，并增加了定量目测和结果确认的价值。所要求的工具将使代表NIH资助的77笔赠款的30名调查人员能够进入全新和有用的调查领域。目前，麻省理工学院没有类似的仪器，合作伙伴医疗保健/哈佛大学或周围地理区域的任何地方都没有。将该仪器放置在首席研究人员的实验室将使与细胞生物学、免疫学、自身免疫学、传染病、肿瘤学和干细胞研究有关的各种研究受益。 2010年2月，在首席调查员的实验室放置了一个ImageStreamX演示单元，为期一周；提交的这5天的试点数据以及2011年重新提交的更新数据。该应用程序中的联合调查员代表了位于MGH的NIH资助的超过2700万美元的工作；整个医院约有1000名首席调查员，NIH的预算约为3.25亿美元。

DESCRIPTION (provided by applicant): NIH NCRR funds are requested in this resubmission to purchase a 5-laser 2-camera Amnis ImageStreamX with three (20x, 40x, 60x) objective lenses. This new platform is a vastly improved, refined and far more powerful version of the original instrument Amnis introduced approximately eleven years ago. This institutionally supported and expertly supervised instrument will be placed in the principal investigator's existing Department of Pathology Flow Cytometry Core which has been in successful operation for almost 30 years. The requested instrument will serve to fill the gap between the highly detailed (but far more slowly acquired) cellular images obtained by confocal microscopy and the imageless but rapidly acquired data obtained by immunofluorescence-based multiparameter flow cytometry. The ImageStreamX combines these two later technologies, providing a synergistic advantage over each used in tandem. The requested instrument quantifies both the intensity and intracellular location of fluorescent cellular probes and can image at high rates of speed (up to ~2000 cells/sec), allowing our consortium of Massachusetts General Hospital (MGH) investigators to analyze rare sub- populations and highly heterogeneous samples with statistically robust and objective results. This technology will permit some investigations that would otherwise be impossible to perform, to actually be implemented. By combining the speed, sensitivity, and immunophenotyping abilities and capacities of flow cytometry with the detailed imagery and functional insights of microscopy, the requested instrument will be an invaluable addition to represented researchers at both downtown Boston and Charlestown research campuses of MGH, and throughout the greater Partners Health Care System. The instrument utilizes the same dyes and markers employed in fluorescence microscopy and flow cytometry and can perform virtually any standard flow cytometry assay with the added value of quantitative visual inspection and resulting confirmation. The requested instrument will enable 30 investigators representing 77 NIH funded grants to enter into entirely new and useful areas of investigation. There is currently no similar instrumentation at MGH, anywhere in Partners Health Care/Harvard University, or the surrounding geographic region. A wide variety of studies related to cell biology, immunology, autoimmune, infectious disease, oncology and stem cell research will benefit from the placement of this instrumentation in the principal investigator's laboratory. An ImageStreamX demo-unit was placed in the principal investigator's laboratory in February 2010 for one week; pilot data produced from those 5-days is presented in this submission as well as updated data for this 2011 re-submission. The co-investigators in this application represent over 27 million dollars of NIH funded work based at MGH; the entire hospital of approximately 1000 principal investigator's has a NIH budget of approximately $325 million dollars.