Biofluid-accessible extracellular vesicles (EVs) may represent a new means to improve the sensitivity and specificity of detecting disease. However, current methods to isolate EVs encounter challenges when they are used to select specific populations. Moreover, it has been difficult to comprehensively characterize heterogeneous EV populations at the single vesicle level. Here, we robustly assessed heterogeneous EV populations from cultured cell lines via nanoparticle tracking analysis, proteomics, transcriptomics, transmission electron microscopy, and quantitative single molecule localization microscopy (qSMLM). Using qSMLM, we quantified the size and biomarker content of individual EVs. We applied qSMLM to patient plasma samples and identified a pancreatic cancer-enriched EV population. Our goal is to advance single molecule characterization of EVs for early disease detection.

Abbreviations: EV: Extracellular Vesicle; qSMLM: quantitative Single Molecule Localization Microscopy; PDAC: Pancreatic Ductal Adenocarcinoma; EGFR: epidermal growth factor receptor 1; CA19-9: carbohydrate antigen 19-9; SEC: size exclusion chromatography; WGA: wheat germ agglutinin; AF647: Alexa Fluor 647; Ab: antibody; HPDEC: Healthy Pancreatic Ductal Epithelial Cell; TEM: Transmission Electron Microscopy.

生物流体可及的细胞外囊泡（EV）可能代表一种新的手段来提高检测疾病的敏感性和特异性。但是，当前用于隔离电动汽车的方法在用于选择特定人群时会遇到挑战。此外，很难在单个囊泡水平上全面表征异种电动汽车群体。在这里，我们通过纳米粒子跟踪分析，蛋白质组学，转录组学，透射电子显微镜和定量单分子定位显微镜（qSMLM），从培养的细胞系中稳健地评估了异种EV群体。使用qSMLM，我们量化了各个电动汽车的大小和生物标志物含量。我们将qSMLM应用于患者血浆样本，并确定了富含胰腺癌的EV人群。

缩写：EV：细胞外囊泡；qSMLM：定量单分子定位显微镜；PDAC：胰腺导管腺癌；EGFR：表皮生长因子受体1；CA19-9：碳水化合物抗原19-9；SEC：尺寸排阻色谱法；WGA：小麦胚芽凝集素；AF647：Alexa Fluor 647；Ab：抗体；HPDEC：健康的胰导管上皮细胞；TEM：透射电子显微镜。