Laser scanning cytometer for high throughput screening

用于高通量筛选的激光扫描细胞仪

• 批准号: 7792645
• 负责人: CAROLINE E SHAMU
• 金额: $ 40.23万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-04 至 2011-03-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792645
• 关键词: Address; Algorithms; Area; Biological; Biological Assay; Biological Process; Boston; Cancer Biology; Cell Death; Cells; Communicable Diseases; Communities; Complex; Data; Development; Devices; Fluorescence; Funding; Generations; Genome; Head; Hospitals; Image; Image Analysis; Lasers; Location; Methods; Microscope; Molecular; Molecular Probes; Neurobiology; Optics; Pathway interactions; RNA Interference; Reader; Recruitment Activity; Relative (related person); Research; Resolution; Resources; Running; Sampling; Scanning; Screening procedure; Site; Technology; Therapeutic; Time; Tube; Whole Organism; Zebrafish; gene function; high throughput screening; instrument; light scattering; medical schools; member; photomultiplier; small molecule; small molecule libraries; therapeutic target; tool

DESCRIPTION (provided by applicant): High-throughput screening (HTS) of small molecule libraries is a widely recognized and increasingly important method for identifying compounds that modulate biological processes. Small molecule compounds thus identified can be powerful molecular probes for dissecting complex biological pathways, and may also be potential leads for development as therapeutics. RNA interference (RNAi) is another powerful tool for probing gene function and, using HTS technologies, can be implemented systematically to screen entire genomes for specific molecular functions or potential therapeutic targets. The ICCB-Longwood Screening Facility (ICCB-L) Research Core provides small molecule and RNAi HTS resources for Harvard Medical School (HMS), HMS-affiliated hospitals, members of the broader Boston-area research community, and other labs nationwide. Here we request funding to purchase a Molecular Devices IsoCyte laser scanning cytometer for use in high-throughput screening at ICCB-L. Although this type of instrument is in use in academic screening centers at other sites in the U.S., no similar instrument currently exists at HMS or in the adjacent research hospitals. As described in this application, the IsoCyte will bring new and much- needed assay capabilities to a large NIH-funded community. The IsoCyte generates low-resolution images of microplate wells. Fluorescence emissions from laser-excited samples are collected using an optical head with a large depth of focus and multiple photomultiplier tubes. A light-scatter mode facilitates generation of images that resemble brightfield images from non-fluorescent samples. Image analysis algorithms run concurrently as images are captured to detect objects (cells) and quantify intensities in up to 4 emission channels. The IsoCyte occupies a useful niche for assay readout between multimode plate readers and screening microscopes. Key features of the IsoCyte relative to plate readers include the greatly enhanced sensitivity of the IsoCyte and the ability to collect single-cell data. The IsoCyte is 5-10 times faster at capturing and analyzing low resolution images than screening microscopes currently in use. Because of its large depth of field, the IsoCyte is ideal for automating imaging of whole organisms such as zebrafish. Such screens are not feasible using conventional screening microscopes. Reflecting the many potential applications of the IsoCyte, this proposal includes specific projects addressing a wide range of topics in the areas of infectious disease, neurobiology, cancer biology, and cell death. Because of its location in the ICCB-L Research Core, the instrument will be well maintained, well managed, and highly accessible to new users as they are recruited.

描述（由申请人提供）：小分子文库的高通量筛选（HTS）是一种广泛认可且日益重要的鉴定调节生物过程的化合物的方法。由此鉴定出的小分子化合物可以成为解析复杂生物途径的强大分子探针，也可能成为治疗药物开发的潜在线索。 RNA 干扰 (RNAi) 是探测基因功能的另一种强大工具，利用 HTS 技术，可以系统地筛选整个基因组的特定分子功能或潜在的治疗靶点。 ICCB-朗伍德筛选设施 (ICCB-L) 研究核心为哈佛医学院 (HMS)、HMS 附属医院、更广泛的波士顿地区研究界成员以及全国其他实验室提供小分子和 RNAi HTS 资源。 在此，我们请求资金购买 Molecular Devices IsoCyte 激光扫描细胞仪，用于 ICCB-L 的高通量筛选。尽管这种类型的仪器已在美国其他地点的学术筛查中心使用，但目前 HMS 或邻近的研究医院还没有类似的仪器。正如本申请中所述，IsoCyte 将为 NIH 资助的大型社区带来急需的新检测功能。 IsoCyte 生成微孔板孔的低分辨率图像。使用具有大焦深和多个光电倍增管的光学头收集激光激发样品的荧光发射。光散射模式有助于生成类似于非荧光样品明场图像的图像。图像分析算法在捕获图像时同时运行，以检测物体（细胞）并量化多达 4 个发射通道的强度。 IsoCyte 在多模式读板器和筛选显微镜之间的检测读数中占有一席之地。 IsoCyte 相对于读板机的主要特点包括 IsoCyte 大大增强的灵敏度和收集单细胞数据的能力。 IsoCyte 捕获和分析低分辨率图像的速度比目前使用的筛选显微镜快 5-10 倍。由于景深较大，IsoCyte 非常适合对斑马鱼等整个生物体进行自动成像。使用传统的筛选显微镜无法进行此类筛选。该提案反映了 IsoCyte 的许多潜在应用，包括解决传染病、神经生物学、癌症生物学和细胞死亡领域广泛主题的具体项目。由于其位于 ICCB-L 研究核心中，该仪器将得到良好的维护、良好的管理，并且在招募新用户时易于使用。