Laser Scanning Cytometer

激光扫描细胞仪

• 批准号: 7595975
• 负责人: Steven A Porcelli
• 金额: $ 39.84万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-22 至 2009-12-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595975
• 关键词: Area; Autoimmune Diseases; Cancer Biology; Cancer Center; Cells; Communicable Diseases; Cultured Cells; Data; Development; Ensure; Faculty; Flow Cytometry; Fluorescence; Funding; Housing; Hybrids; Image; Image Analysis; Immunology; Institution; Label; Laboratories; Lasers; Life; Microscope; Microscopic; Microscopy; Monitor; Operative Surgical Procedures; Request for Applications; Research Personnel; Research Project Grants; Sampling; Scanning; Science; Signal Transduction; Slide; Specimen; Staging; Supervision; Surface; Technology; Time; Tissues; Translational Research; Tube; cell fixing; detector; experience; instrument; lens; meetings; member; new technology; photomultiplier; public health relevance

DESCRIPTION (provided by applicant): We are requesting funds for a Compucyte iCys Laser Scanning Cytometer (LSC). The LSC is a relatively recently developed instrument that joins many of the advantages of traditional microscopy and flow cytometry, and significantly extends the limits of single cell analyses. Rather than randomly imaging an entire field like a microscope does, it selects, images and analyzes cells that meet certain adjustable criteria (such as size or fluorescence). Unlike conventional flow cytometry, the LSC is usually used to analyze cells fixed to horizontal surface, such as cultured cells on slides or in a variety of culture vessels. Live cells can be imaged and cytometrically analyzed for fluorescent signals, and a variety of types of fixed specimens can also be studied in this way. The instrument uses an automated nanostep X-Y stage to move samples through the objective lens, applying up to three different lasers to excite fluorescent emissions and quantitating these using an array of four photomultiplier tube (PMT) detectors. Thus, the LSC is essentially a powerful hybrid technology that combines imaging and cytometric analysis in a single platform. Importantly, with the LSC image becomes a parameter, and detailed microscopic pictures are stored and referenced directly to cytometric data. A highly appealing feature of the instrument that we plan to use extensively is its ability to gather quantitative fluorescence data from microscopic areas of fluorescently labeled tissue sections, and to display and analyze such data in a manner that is comparable in many cases to true cytometric data such as one obtains with traditional flow cytometry. We plan to house and operate the iCys LSC in our well established central Flow Cytometry Core Laboratory, where there will be ample expertise and oversight to ensure that the instrument is used to its full capacity. The Core Laboratory has an experienced senior faculty member (Dr. Steven Porcelli) as its supervisor, and employs three full time faculty level associates to direct operations and provide technical supervision. There is also a range of internal consultants with expertise in areas of major relevance to the development of applications that will use the LSC. The Core Laboratory has a well developed and carefully monitored financial plan, and receives substantial institutional support from the institution and from our NCI-funded Cancer Center to ensure its efficient and stable operation. By incorporating the iCys LSC into this core laboratory, a large body of investigators pursuing many diverse NIH-funded basic and translational research projects will gain convenient and affordable access to this important new technology. Public Health Relevance: This application requests funding that will allow our Flow Cytometry Core Laboratory to acquire a new analytical instrument called the Compucyte iCys Laser Scanning Cytometer. This instrument will provide many researchers at our institution with a powerful new way to analyze cells and tissues. The studies that will be done with this instrument will advance our understanding of a wide spectrum of areas with high relevance to biomedical science, including cancer biology, immunology, autoimmune diseases and infectious diseases.

描述（由申请人提供）：我们正在申请资金购买 Compucyte iCys 激光扫描细胞仪 (LSC)。 LSC 是一种相对较新开发的仪器，结合了传统显微镜和流式细胞术的许多优点，并显着扩展了单细胞分析的极限。它不像显微镜那样对整个视野进行随机成像，而是选择、成像和分析满足某些可调整标准（例如大小或荧光）的细胞。与传统的流式细胞术不同，LSC通常用于分析固定在水平表面的细胞，例如载玻片上或各种培养容器中的培养细胞。可以对活细胞进行成像并通过细胞计数分析荧光信号，也可以通过这种方式研究各种类型的固定样本。该仪器使用自动纳级 X-Y 平台将样品移动通过物镜，应用多达三种不同的激光来激发荧光发射，并使用四个光电倍增管 (PMT) 检测器阵列对这些荧光进行定量。因此，LSC 本质上是一种强大的混合技术，将成像和细胞计数分析结合在一个平台中。重要的是，LSC 图像成为一个参数，详细的显微图片被存储并直接引用到细胞计数数据。我们计划广泛使用的该仪器的一个非常吸引人的功能是它能够从荧光标记的组织切片的微观区域收集定量荧光数据，并以在许多情况下与真实细胞计数数据相当的方式显示和分析这些数据。例如通过传统流式细胞术获得的结果。我们计划在我们完善的中央流式细胞术核心实验室安置和操作 iCys LSC，那里将有充足的专业知识和监督，以确保仪器充分发挥作用。核心实验室拥有经验丰富的高级教员（Steven Porcelli 博士）担任主管，并聘请了三名全职教员级助理来指导操作并提供技术监督。还有一系列内部顾问，他们在与开发将使用 LSC 的应用程序相关的主要领域具有专业知识。核心实验室拥有完善且严格监控的财务计划，并得到该机构和我们 NCI 资助的癌症中心的大力机构支持，以确保其高效稳定的运行。通过将 iCys LSC 纳入这个核心实验室，众多从事 NIH 资助的各种基础和转化研究项目的研究人员将能够方便且经济地获得这项重要的新技术。公共卫生相关性：本申请请求资金，以便我们的流式细胞术核心实验室能够购买一种名为 Compucyte iCys 激光扫描细胞仪的新型分析仪器。该仪器将为我们机构的许多研究人员提供分析细胞和组织的强大新方法。使用该仪器进行的研究将增进我们对与生物医学高度相关的广泛领域的理解，包括癌症生物学、免疫学、自身免疫性疾病和传染病。