ImageStream X Mark II for NJMS Flow Core

描述(由申请人提供)：新泽西医学院这份共享仪器补助金(SIG)申请的调查人员申请资金购买最先进的Amennis ImageStreamX Mark II成像流式细胞仪，以取代我们于2007年底购买的过时的第一代ImageStream 100，并将SIG授予当前申请的PI。ImageStreamX Mark II将通过NJMS的流式细胞术和免疫学核心实验室进行容纳和管理，该实验室得到了NJMS的强有力的机构支持。成像流式细胞术提供了传统流式细胞术的高通量内容，并能够对流动中的细胞群体进行图像分析。这项技术允许分析，例如，核移位、共定位研究、细胞内病原体跟踪、吞噬、FRET和细胞群中的凋亡。我们要求ImageStreamX Mark II配备四个激励激光器：488、405、561和592 nm激光器，外加标准的785 nm SSC激光器。此外，我们要求该仪器配备12个成像通道、可提供灵活的20X、40X和60X成像能力的多料盒选项、扩展景深元素和96孔板自动取样器。最后，我们需要一个独立的工作站来进行数据分析。申请的ImageStreamX Mark II比第一代仪器有多项改进：最重要的是，应要求的ImageStreamX Mark II将允许在单个样本中使用多达10个荧光体，并且比最初的ImageStream 100快20倍。与我们原始仪器的四种颜色相比，同时评估成像流中的10个标志物的能力大大扩展了仪器的能力，并允许研究人员将用于传统流动和细胞分选的多色流动面板应用于ImageStreamX Mark II。这些对仪器的改进，以及多MAG元素，使ImageStreamX Mark II成为一种更多功能的仪器，可用于进行复杂的多颜色流式细胞仪分析，即使是对非常罕见的细胞也是如此。SIG的调查人员代表了一群来自新泽西医学院不同部门的高生产率科学家，他们几乎都有NIH R01的资助。SIG的许多研究人员已经有效地利用了我们现有的ImageStream 100，并发表了论文并获得了使用ImageStream数据的资助，但发现需要更复杂的成像流式细胞术。此SIG应用程序有8个主要用户和5个主要用户。虽然许多人是免疫学家，但SIG的调查人员还包括一名神经学家、一名细胞生理学家、一名微生物学家和生物化学家。因此，ImageStreamX Mark II有望在多种学科中提供有价值的成像流式细胞仪功能。这一工具将提高SIG研究人员的生产力，也将为博士后和博士后实习生提供宝贵的机会。

DESCRIPTION (provided by applicant): The investigators in this Shared Instrument Grant (SIG) application at New Jersey Medical School request funds for purchase of a state-of-the-art Amnis ImageStreamX Mark II imaging flow cytometer to replace our outdated, first-generation ImageStream 100 that was purchased in late 2007 with an SIG awarded to the PI of the current application. The ImageStreamX Mark II will be housed and administrated through the Flow Cytometry and Immunology Core Laboratory at NJMS, which has strong institutional support from NJMS. Imaging flow cytometry provides the high-throughput content of traditional flow cytometry coupled with the ability to carry-out image analysis on cell populations in flow. This technology allows for analysis, for example, of nuclear translocation, co-localization studies, intracellular pathogen tracking, phagocytosis, FRET, and apoptosis in populations of cells. We are requesting that the ImageStreamX Mark II be equipped with four excitation lasers: 488, 405, 561 and 592nm lasers, plus the standard 785nm SSC laser. In addition, we request that the instrument be equipped with 12 imaging channels, the Multi-Mag option to provide flexible 20X, 40X and 60X imaging capabilities, an extended depth-of-field element, and a 96-well plate auto-sampler. Finally, we request a stand-alone work station for data analysis. The requested ImageStreamX Mark II has multiple improvements over the first generation instrument: most importantly, the ImageStreamX Mark II as requested will allow for up to 10-fluorochomes to be used in a single sample and is up to 20X faster than the original ImageStream 100. The ability to simultaneously evaluate 10 markers in imaging flow dramatically extends the capability of the instrument compared to the four colors with our original instrument and allows investigators to adapt multi-color flow panels used for traditional flow and cell sorting to the ImageStreamX Mark II. These improvements to the instrumentation, along with the Multi-Mag element make the ImageStreamX Mark II a much more versatile instrument that can be used to carry-out sophisticated, multi-color flow cytometric analysis, even on very rare cells. The SIG investigators represent a group of highly-productive scientists from diverse departments at the New Jersey Medical School, almost all of whom have NIH R01 funding. Many of the SIG investigators have effectively utilized our existing ImageStream 100 and have published papers and obtained grants with ImageStream data, but have found the need for more sophisticated imaging flow cytometry. There are 8 major and 5 major users for this SIG application. While many are immunologists, the SIG investigators also include a neuroscientist, a cell physiologist, a microbiologist and biochemists. Thus, the ImageStreamX Mark II is expected to provide valuable imaging flow cytometry capabilities across a wide diversity of disciplines. This instrument will enhance the productivity of the SIG investigators and will also provide valuable opportunities for pre- and post-doctoral trainees.