Assay Dev/Adapt (California)

检测开发/适应（加利福尼亚州）

• 批准号: 8538721
• 负责人: Steven J Brown
• 金额: $ 94.88万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8538721
• 关键词: Adherence; Applications Grants; Automation; Biological Assay; Business Rules; Calcium; California; Cataloging; Catalogs; Cell Count; Cell Line; Cells; Cellular biology; Chemicals; Chemistry; Collaborations; Color; Computer software; Critical Pathways; Cyclic AMP; Data; Data Files; Detection; Development; Development Plans; Dimethyl Sulfoxide; Dose; Ensure; Enzymes; Equipment; Fluorescein-5-isothiocyanate; Fluorescence; Fluorescence Microscopy; Follow-Up Studies; Freezing; G Protein-Coupled Receptor Genes; Generic Drugs; Hand; Hour; Housing; Image; Image Analysis; Incubators; Label; Laboratories; Lactamase; Lasers; Length; Life; Measurement; Measures; Microinjections; Microscope; Microscopy; Molecular Biology; Mycoplasma; Nucleic Acids; Outsourcing; Peptides; Performance; Process; Production; Proteins; Protocols documentation; Provider; Readiness; Reagent; Reporter; Reporting; Research Personnel; Resources; Running; Sampling; Scanning; Schedule; Science; Screening procedure; Secure; Source; Specificity; Stream; Surveys; System; Teleconferences; Temperature; Testing; Time; TimeLine; Titrations; United States National Institutes of Health; Vendor; Vial device; Visual; Work; assay development; base; continuous cell line; counterscreen; enzyme substrate; fluorescence microscope; follow-up; foot; inhibitor/antagonist; meetings; micromanipulator; release of sequestered calcium ion into cytoplasm; response; scale up; tissue culture

ASSAY DEVELOPMENT/ADAPTATION CORE In La Jolla, 600 square ft of laboratory space is dedicated to the MLPCN effort. The lab is configured for maximal use of space with the tissue culture room adjacent to the automation bench. The tissue culture room has two 6-foot biosafety cabinets. In addition, the PI has 2500 sq. ft lab space immediately below the assay development facility. The lab is fully equipped for cell and molecular biology and is used in assay development efforts requiring these capabilities. The tissue culture lab has a Zeiss Axiovert fluorescent microscope equipped with filters for visual detection of green fluorescence (FITC, GFP) and CCF2/4 (b-lactamase substrate) loaded cells. The ICND building has a core microscopy facility available for secondary assays (e.g. GPCR internalization) with the following equipment. ¿ Brightfield and fluorescence microscopes ¿ Dissecting stereoscope with color camera ¿ Calcium imaging automated system with micromanipulator for cell picking Olympus confocal laser-scanning microscope with 4 laser lines ¿ DeltaVision deconvolution acquisition system ¿ Spinning-disk confocal microscope (2 laser lines) with incubator chamber for live-cell fluorescence microscopy ¿ Live-cell brightfield microscope with incubator ¿ Eppendorf microinjection system on inverted fluorescence microscope ¿ MetaMorph 6.5 acquisition and image analysis software

试验开发/适应性核心 在拉霍亚，600平方英尺的实验室空间专门用于MLPCN的工作。实验室配置为 最大限度地利用空间，组织培养室与自动化工作台相邻。组织培养室 有两个六英尺高的生物安全柜 此外，PI有2500平方米。位于测定开发设施正下方的30英尺实验室空间。实验室完全 配备细胞和分子生物学，并用于需要这些能力的测定开发工作。 组织培养实验室有一台蔡司Axiovert荧光显微镜，配有滤光片，用于目视检测 绿色荧光（FITC，GFP）和CCF 2/4（β-内酰胺酶底物）加载的细胞。 ICND大楼有一个核心显微镜设施，可用于二次检测（例如GPCR内化） 以下设备。 荧光显微镜和荧光显微镜 带彩色摄像机的解剖立体镜 <$钙成像自动化系统与显微操作器的细胞挑选 Olympus共聚焦激光扫描显微镜，带4条激光线 DeltaVision反卷积采集系统 Ø转盘共聚焦显微镜（2条激光线），带活细胞荧光培养箱 显微镜 带培养箱的活细胞明视野显微镜 倒置荧光显微镜上的Eppendorf显微注射系统 MetaMorph 6.5采集和图像分析软件