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Protein Profiling and Sizing of Extracellular Vesicles from Colorectal Cancer Patients via Flow Cytometry
ACS Nano ( IF 15.8 ) Pub Date : 2018-01-08 00:00:00 , DOI: 10.1021/acsnano.7b07782 Ye Tian 1 , Ling Ma 1 , Manfei Gong 1 , Guoqiang Su 2 , Shaobin Zhu 1 , Wenqiang Zhang 1 , Shuo Wang 1 , Zhibin Li 3 , Chaoxiang Chen 1 , Lihong Li 1 , Lina Wu 1 , Xiaomei Yan 1
ACS Nano ( IF 15.8 ) Pub Date : 2018-01-08 00:00:00 , DOI: 10.1021/acsnano.7b07782 Ye Tian 1 , Ling Ma 1 , Manfei Gong 1 , Guoqiang Su 2 , Shaobin Zhu 1 , Wenqiang Zhang 1 , Shuo Wang 1 , Zhibin Li 3 , Chaoxiang Chen 1 , Lihong Li 1 , Lina Wu 1 , Xiaomei Yan 1
Affiliation
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Extracellular vesicles (EVs) have stimulated considerable scientific and clinical interest, yet protein profiling and sizing of individual EVs remains challenging due to their small particle size, low abundance of proteins, and overall heterogeneity. Building upon a laboratory-built high-sensitivity flow cytometer (HSFCM), we report here a rapid approach for quantitative multiparameter analysis of single EVs down to 40 nm with an analysis rate up to 10 000 particles per minute. Statistically robust particle size distribution was acquired in minutes with a resolution and profile well matched with those of cryo-TEM measurements. Subpopulations of EVs expressing CD9, CD63, and/or CD81 were quantified upon immunofluorescent staining. When HSFCM was used to analyze blood samples, a significantly elevated level of CD147-positive EVs was identified in colorectal cancer patients compared to healthy controls (P < 0.001). HSFCM provides a sensitive and rapid platform for surface protein profiling and sizing of individual EVs, which could greatly aid the understanding of EV-mediated intercellular communication and the development of advanced diagnostic and therapeutic strategies.
中文翻译：
蛋白质谱和结直肠癌患者外囊的大小调整通过流式细胞仪
细胞外囊泡（EV）引起了相当大的科学和临床兴趣，但是由于其体积小，蛋白质丰度低和整体异质性，单个EV的蛋白质谱分析和尺寸测定仍然具有挑战性。在实验室建造的高灵敏度流式细胞仪（HSFCM）的基础上，我们在此报告了一种快速的方法，可以对低至40 nm的单个EV进行定量多参数分析，分析速率最高为每分钟10000个颗粒。数分钟内就获得了统计上稳定的粒度分布，其分辨率和轮廓与冷冻TEM测量的分辨率和轮廓非常匹配。在免疫荧光染色后，对表达CD9，CD63和/或CD81的EV的亚群进行定量。当使用HSFCM分析血液样本时，P <0.001）。HSFCM为单个电动汽车的表面蛋白分析和定型提供了一个灵敏而快速的平台，这可以极大地帮助理解电动汽车介导的细胞间通讯以及发展先进的诊断和治疗策略。
更新日期：2018-01-08
中文翻译：
蛋白质谱和结直肠癌患者外囊的大小调整通过流式细胞仪
细胞外囊泡（EV）引起了相当大的科学和临床兴趣，但是由于其体积小，蛋白质丰度低和整体异质性，单个EV的蛋白质谱分析和尺寸测定仍然具有挑战性。在实验室建造的高灵敏度流式细胞仪（HSFCM）的基础上，我们在此报告了一种快速的方法，可以对低至40 nm的单个EV进行定量多参数分析，分析速率最高为每分钟10000个颗粒。数分钟内就获得了统计上稳定的粒度分布，其分辨率和轮廓与冷冻TEM测量的分辨率和轮廓非常匹配。在免疫荧光染色后，对表达CD9，CD63和/或CD81的EV的亚群进行定量。当使用HSFCM分析血液样本时，P <0.001）。HSFCM为单个电动汽车的表面蛋白分析和定型提供了一个灵敏而快速的平台，这可以极大地帮助理解电动汽车介导的细胞间通讯以及发展先进的诊断和治疗策略。
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