化学
荧光
共焦
荧光显微镜
单细胞分析
细胞计数
癌细胞
共焦显微镜
纳米技术
显微镜
细胞仪
细胞
癌症
材料科学
光学
细胞周期
生物化学
物理
医学
内科学
作者
Shengkai Zhang,Lifang Yao,Xiaofei Lv,Meijun Lu,Fengli Gao,Liangliang Zhang,Shulin Zhao,Shengqiang Hu
标识
DOI:10.1021/acs.analchem.2c05684
摘要
Fluorescence-assisted digital counting analysis allowed sensitive quantification of targets by measuring individual fluorescent labels. However, traditional fluorescent labels suffered from low brightness, small size, and sophisticated preparation procedures. Herein, engineering fluorescent dye-stained cancer cells with magnetic nanoparticles were proposed to construct single-cell probes for fluorescence-assisted digital counting analysis by quantifying the target-dependent binding or cleaving events. Various engineering strategies of cancer cells including biological recognition and chemical modification were developed for rationally designing single-cell probes. Introduction of suitable recognition elements into single-cell probes allowed digital quantification of each target-dependent event via counting the colored single-cell probes in the representative image taken using a confocal microscope. The reliability of the proposed digital counting strategy was corroborated by traditional optical microscopy- and flow cytometry-dependent counting technologies. The advantages of single-cell probes, including high brightness, big size, ease of preparation, and magnetic separation, contributed to the sensitive and selective analysis of targets of interest. As proof-to-concept assays, indirect analysis of exonuclease III (Exo III) activity, as well as direct quantitation of cancer cells, were investigated, and the potential in biological sample analysis was also assessed. This sensing strategy will open a new avenue for the development of biosensors.
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