Department of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Anal Chem. 2021 Oct 5;93(39):13152-13160. doi: 10.1021/acs.analchem.1c01446. Epub 2021 Sep 22.
Single-cell EV (extracellular vesicle) secretion analysis is emerging for a better understanding of non-genetic cellular heterogeneity regulating human health and diseases through intercellular mediators. However, the requirements of expensive and bulky instrumentations hinder its widespread use. Herein, by combining gold nanoparticle-enhanced silver staining and the Poisson distribution, we reported the use of a home-use scanner to realize high-throughput single-cell EV secretion analysis without cell counting. We applied the platform to analyze the secretions of different EV phenotypes with the human oral squamous cell carcinoma cell line and primary cells from patients, which generated single-cell results comparable with those of the immunofluorescence approach. Notably, we also realized the quantification of the number of EVs secreted from every single cell using their respective titration curves obtained from population samples, making it possible to directly compare different EV phonotypes in regard to their secretion number, secretion rate, and so forth. The technology introduced here is simple, easy to operate, and of low cost, which make it a potential, easily accessible, and affordable tool for widespread use in both basic and clinical research.
单细胞 EV(细胞外囊泡)分泌分析正在兴起,通过细胞间介质,可以更好地了解调节人类健康和疾病的非遗传细胞异质性。然而,昂贵且庞大的仪器设备的要求阻碍了其广泛应用。在此,我们通过结合金纳米粒子增强的银染色和泊松分布,报告了使用家用扫描仪实现高通量单细胞 EV 分泌分析而无需细胞计数的方法。我们应用该平台分析了不同 EV 表型的分泌情况,使用人口腔鳞状细胞癌细胞系和来自患者的原代细胞,得到的单细胞结果与免疫荧光方法相当。值得注意的是,我们还使用从群体样本中获得的各自滴定曲线,实现了从每个单细胞分泌的 EV 数量的定量,使得可以直接比较不同 EV 表型的分泌数量、分泌率等。这里介绍的技术简单、易于操作且成本低,使其成为一种具有潜力、易于获取且负担得起的工具,可广泛应用于基础和临床研究。