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High-Throughput Single-Cell Extracellular Vesicle Secretion Analysis on a Desktop Scanner without Cell Counting
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-09-22 , DOI: 10.1021/acs.analchem.1c01446 Fengjiao Zhu 1, 2 , Yahui Ji 1 , Linmei Li 1 , Xue Bai 1 , Xianming Liu 1 , Yong Luo 3 , Tingjiao Liu 4 , Bingcheng Lin 1 , Yao Lu 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-09-22 , DOI: 10.1021/acs.analchem.1c01446 Fengjiao Zhu 1, 2 , Yahui Ji 1 , Linmei Li 1 , Xue Bai 1 , Xianming Liu 1 , Yong Luo 3 , Tingjiao Liu 4 , Bingcheng Lin 1 , Yao Lu 1
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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.
中文翻译：
在台式扫描仪上进行高通量单细胞细胞外囊泡分泌分析，无需细胞计数
单细胞 EV（细胞外囊泡）分泌分析正在兴起，以更好地了解通过细胞间介质调节人类健康和疾病的非遗传细胞异质性。然而，昂贵且笨重的仪器的要求阻碍了其广泛使用。在此，通过结合金纳米粒子增强银染色和泊松分布，我们报道了使用家用扫描仪实现高通量单细胞 EV 分泌分析，无需细胞计数。我们应用该平台用人口腔鳞状细胞癌细胞系和患者的原代细胞分析不同 EV 表型的分泌物，产生与免疫荧光方法相当的单细胞结果。尤其，我们还使用从群体样本中获得的各自滴定曲线实现了每个细胞分泌的 EV 数量的量化，从而可以直接比较不同 EV 音型的分泌数量、分泌速率等。这里介绍的技术简单、易于操作且成本低，使其成为一种潜在、易于获取且价格合理的工具，可广泛用于基础和临床研究。
更新日期：2021-10-06
中文翻译：
在台式扫描仪上进行高通量单细胞细胞外囊泡分泌分析，无需细胞计数
单细胞 EV（细胞外囊泡）分泌分析正在兴起，以更好地了解通过细胞间介质调节人类健康和疾病的非遗传细胞异质性。然而，昂贵且笨重的仪器的要求阻碍了其广泛使用。在此，通过结合金纳米粒子增强银染色和泊松分布，我们报道了使用家用扫描仪实现高通量单细胞 EV 分泌分析，无需细胞计数。我们应用该平台用人口腔鳞状细胞癌细胞系和患者的原代细胞分析不同 EV 表型的分泌物，产生与免疫荧光方法相当的单细胞结果。尤其，我们还使用从群体样本中获得的各自滴定曲线实现了每个细胞分泌的 EV 数量的量化，从而可以直接比较不同 EV 音型的分泌数量、分泌速率等。这里介绍的技术简单、易于操作且成本低，使其成为一种潜在、易于获取且价格合理的工具，可广泛用于基础和临床研究。
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