State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, PR China.
Anal Chem. 2012 Nov 20;84(22):10097-104. doi: 10.1021/ac302609y. Epub 2012 Nov 1.
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.
从稀有细胞样本中获取丰富的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。然而，这种类型的数据通常受到传统分析技术（如流式细胞术）的限制。本文中，我们描述了一种与基于量子点的（QDs）免疫荧光（IF）方法相结合的微流控平台，用于测量单细胞或细胞群体表面糖链的表达。与传统的 IF 染色相比，基于 QDs 的 IF 探针表现出更高的亮度和抗光漂白稳定性。利用新型 IF 染色方案和微流控平台的优点，可进行高通量 IF 染色以测量单个 K562 细胞在药物处理后的糖链表达和变化。由于间接 IF 染色中信号的放大，该方案在糖链表达谱方面具有较高的灵敏度。细胞样本的大小仅为 4×10(3)个细胞，这使得稀有细胞样本的分析变得可行。该方法可能会广泛应用于评估细胞表面糖蛋白的表达以及以高通量方式分析细胞群体的异质性。
