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限制。它不像显微镜那样随机地对整个视野进行成像，而是选择、成像和分析满足某些可调节标准（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上的细胞，例如载玻片上或各种培养容器中的培养细胞。活细胞可以成像和细胞计数分析荧光信号，各种类型的固定标本也可以通过这种方式进行研究。该仪器使用自动纳米级X-Y载物台将样品移动通过物镜透镜，应用多达三种不同的激光激发荧光发射，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上。重要的是，LSC图像成为一个参数，详细的显微照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仪器的一个非常有吸引力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正的细胞计数数据（如用传统流式细胞术获得的数据）相当的方式显示和分析这些数据。我们计划在我们完善的中央流式细胞术核心实验室中安置和操作iCys LSC，在那里将有充足的专业知识和监督，以确保仪器充分利用其能力。核心实验室有一位经验丰富的高级教师（Steven Porcelli博士）作为其主管，并雇用了三名全职教师级别的同事来指导操作和提供技术监督。还有一系列内部顾问，他们在与使用LSC的应用程序开发相关的领域具有专业知识。核心实验室有一个完善的和仔细监测的财务计划，并从该机构和我们的NCI资助的癌症中心获得大量的机构支持，以确保其有效和稳定的运作。通过将iCys LSC纳入这个核心实验室，大量从事NIH资助的基础和转化研究项目的研究人员将方便且负担得起地获得这项重要的新技术。公共卫生相关性：该申请要求提供资金，使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分析仪器。该仪器将为我们机构的许多研究人员提供一种分析细胞和组织的强大新方法。将使用该仪器进行的研究将促进我们对与生物医学科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。