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基于光学成像光纤的单个活细胞阵列:一种高密度细胞检测平台。

Optical imaging fiber-based single live cell arrays: a high-density cell assay platform.

作者信息

Biran Israel, Walt David R

机构信息

Department of Chemistry, Tufts University, Medford, Massachusetts 02155, USA.

出版信息

Anal Chem. 2002 Jul 1;74(13):3046-54. doi: 10.1021/ac020009e.

Abstract

A high-density, ordered array containing thousands of microwells is fabricated on an optical imaging fiber. Each individually addressable microwell is used to accommodate a single living cell. A charged coupled device (CCD) detector is employed to monitor and spatially resolve the fluorescence signals obtained from each individual cell, allowing simultaneous monitoring of cellular responses of all the cells in the array using reporter genes (lacZ, EGFP, ECFP, DsRed) or fluorescent indicators. Yeast and bacteria cell arrays were fabricated and used to perform multiplexed cell assays with resolution at the single-cell level. Monitoring gene expression in single yeast cells carrying a two-hybrid system was used to detect in vivo protein-protein interactions. The single-cell array technology provides a new platform for monitoring the unique multiple responses of large populations of individual cells from different strains or cell lines. The rich data acquired by the cell array has the potential to be employed as a new tool for cell biology research as well as to improve cell-based high-throughput screening (HTS) applications, such as the validation of new disease-associated cellular targets and the early-stage evaluation of potential drug candidates.

摘要

在一根光学成像光纤上制造出一个包含数千个微孔的高密度、有序阵列。每个可单独寻址的微孔用于容纳单个活细胞。采用电荷耦合器件(CCD)探测器来监测并在空间上分辨从每个单个细胞获得的荧光信号,从而能够使用报告基因(lacZ、增强型绿色荧光蛋白(EGFP)、增强型青色荧光蛋白(ECFP)、红色荧光蛋白(DsRed))或荧光指示剂同时监测阵列中所有细胞的细胞反应。制造了酵母和细菌细胞阵列,并用于进行单细胞水平分辨率的多重细胞分析。监测携带双杂交系统的单个酵母细胞中的基因表达,以检测体内蛋白质 - 蛋白质相互作用。单细胞阵列技术为监测来自不同菌株或细胞系的大量单个细胞的独特多种反应提供了一个新平台。细胞阵列获取的丰富数据有潜力用作细胞生物学研究的新工具,以及改进基于细胞的高通量筛选(HTS)应用,例如新的疾病相关细胞靶点的验证和潜在药物候选物的早期评估。

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