Live-cell Microarray for high-throughput observation of metabolic signatures

用于高通量观察代谢特征的活细胞微阵列

• 批准号: 8333992
• 负责人: Deirdre R. Meldrum
• 金额: $ 72.54万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-16 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8333992
• 关键词: Address; Biological; Cells; Chemicals; Collaborations; Data; Data Set; Databases; Detection; Development; Device or Instrument Development; Devices; Disease; Effectiveness; Ensure; Funding; Generations; Glass; Glucose; Health; Heterogeneity; Human; Human Pathology; Incubated; Individual; Instruction; Label; Laboratories; Lasers; Libraries; Life; Malignant Neoplasms; Measurement; Measures; Metabolic; Metabolism; Metals; Methods; Molecular; Monitor; Network-based; Noise; Oxygen; Oxygen Consumption; Pathway interactions; Phenotype; Polymers; Population; Procedures; Process; Reading; Resolution; Running; Sampling; Scanning; Screening procedure; Signal Transduction; Speed; Supporting Cell; System; Technology; Translations; Writing; base; biosignature; cell type; extracellular; glucose uptake; insight; instrument; programs; response; seal; sensor; single cell analysis; time interval; tool

High throughput live-cell microarray screening technology for dynamic, multiparameter sensing cf single-cell metabolic phenotypes is proposed. The proposal addresses Common Fund priorities by extending the range of signatures available to the LINCS centers. A sandwich microarray, called the "Cellarium" will be developed and used to analyze individual live cells. The bottom layer ofthe sandwich supports cells in shallow microwells etched in glass. The top layer ofthe sandwich seals the cells in the 150-picoliter microwells, and incorporates extracellular fluorescent sensors for multiparameter detection ofthe metabolic analytes, oxygen, pH and glucose. Chemical isolation is achieved when the two layers are compressed together with a flat metal spring allowing dynamic measurement of transmembrane fluxes without Intracellular probes. Single-cell analysis which directly reveals heterogeneity in metabolic response to perturbations within an isogenic cell population is critical to biological inference. This microarray is an extensible tool for deriving a standardized multiparameter set of data that can be integrated in a coordinated way into LINCS. The specific aims ofthe project are: 1) develop a disposable microarray ("Cellarium") for dynamic, high throughput, multiparameter metabolic measurements of perturbation-induced signatures of live single cells; 2) modify a commercial microarray scanner to read out the Cellarium; 3) verify the effectiveness of this technology across a range of cell types by simultaneously monitoring 02, pH, glucose and ATP responses; 4) validate the platform by analyzing the distribution of metabolic signatures of single cells in response to perturbations; 5) develop written and graphical standard operating procedures that enable reproducible data generation; 6) develop active participation with LINCS partners in the instrument development process to ensure efficient device and methods translation.

用于单细胞动态多参数传感的高通量活细胞微阵列筛选技术