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Quantum Dots-Based Immunofluorescent Microfluidic Chip for the Analysis of Glycan Expression at Single-Cells

机译:基于量子点的免疫荧光微流控芯片,用于分析单细胞中聚糖的表达

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摘要

Rich high-quality single-cell information from rare cell sample is very important for the quantitative systems biology description of cellular function. However, this type of data is often prohibited by the conventional analytical technology such as flow cytometry. In this paper, we described a microfluidic platform coupled with a quantum dots-based (QDs) immunofluorescence (IF) approach to measure the expression of glycans on the cell surface of single cells or cell population. Compared with conventional IF staining, the QDs-based IF probe exhibited higher brightness and stability against photobleaching. With the merits of the novel IF staining protocol and microfluidic platform, high-throughput IF staining was performed to measure the glycan expressions and the changes at single K562 cells after drug treatment. The protocol proposed here showed a high sensitivity on the glycan expression profile owing to the amplification of the signal in indirect IF staining. The size of cell sample was only 4 × 10~3 cells, which made the rare cell sample analysis accessible. This method may find widespread application for assessing cell-surface glycoprotein expression as well as analysis of the heterogeneity in cell populations in a high-throughput manner.
机译:来自稀有细胞样品的丰富高质量单细胞信息对于细胞功能的定量系统生物学描述非常重要。但是,此类数据通常被常规分析技术(例如流式细胞仪)所禁止。在本文中,我们描述了一种微流体平台,结合基于量子点(QDs)的免疫荧光(IF)方法来测量单细胞或细胞群体的细胞表面上聚糖的表达。与传统的IF染色相比,基于QDs的IF探针具有更高的亮度和抗光漂白的稳定性。利用新型IF染色方案和微流体平台的优点,进行了高通量IF染色,以检测药物处理后单个K562细胞的聚糖表达和变化。由于间接IF染色中信号的放大,此处提出的方案对聚糖表达谱表现出高度的敏感性。细胞样品的大小仅为4×10〜3个细胞,这使得稀有细胞样品分析变得容易进行。该方法可广泛用于评估细胞表面糖蛋白表达以及以高通量方式分析细胞群体中的异质性。

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