Assay Dev/Adapt (California)

检测开发/适应（加利福尼亚州）

• 批准号: 8525451
• 负责人: Steven J Brown
• 金额: $ 179.59万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8525451
• 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; 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; screening; 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

测定开发/适应核心 在拉霍亚（La Jolla），实验室空间的600平方英尺致力于MLPCN的工作。实验室配置为 与自动化台相邻的组织培养室的空间最大使用。组织培养室 有两个6英尺的生物安全柜。 此外，PI在评估开发设施下方有2500平方英尺的实验室空间。实验室完全 配备了细胞和分子生物学，用于测定需要这些能力的开发工​​作。 组织培养实验室具有配备过滤器的Zeiss Axiovert荧光显微镜，可视检测 绿色荧光（FITC，GFP）和CCF2/4（B-内乳糖酶底物）负载的细胞。 ICND建筑具有可用于辅助测定的核心显微镜设施（例如GPCR内在化） 使用以下设备。 �Brightfield和荧光显微镜 »用彩色摄像头解剖立体镜 �钙成像自动化系统，带有微型操纵器用于细胞采摘 奥林巴斯共聚焦激光扫描显微镜与4个激光线 »deltavision反卷积采集系统 »与孵化室的旋转盘共聚焦显微镜（2个激光线），用于活细胞荧光 显微镜 »带有孵化器的活细胞Brightfield显微镜 »倒荧光显微镜上的Eppendorf显微注射系统 „ Metamorph 6.5获取和图像分析软件