Membrane vesicles (MVs) play biologically important roles in Gram-positive bacteria, and purification is essential for their study. Although high-performance flow cytometry has the capability to quantify and isolate specific small particles, it has not been examined for MV isolation. In this study, we used high-performance flow cytometry to analyze MV from Gram-positive bacteria, Staphylococcus aureus and Bacillus subtilis, prepared by iodixanol density-gradient ultracentrifugation. Analysis of the quality of MV samples before and after sorting showed that the flow cytometric sorting provided higher purity and uniformity compared to gradient isolation alone. The MV purification method using flow cytometry should prove useful for applications requiring a very high purity of MV samples such as proteomic, metagenomic or lipidomic studies.

膜囊泡 (MV) 在革兰氏阳性菌中发挥着重要的生物学作用，纯化对其研究至关重要。尽管高性能流式细胞术具有定量和分离特定小颗粒的能力，但尚未对其进行 MV 分离检查。在本研究中，我们使用高性能流式细胞术分析了革兰氏阳性菌、金黄色葡萄球菌和枯草芽孢杆菌的MV ，碘克沙醇密度梯度超速离心法制备。分选前后 MV 样品的质量分析表明，与单独的梯度分离相比，流式细胞仪分选提供了更高的纯度和均匀性。使用流式细胞术的 MV 纯化方法应证明适用于需要非常高纯度的 MV 样品的应用，例如蛋白质组学、宏基因组学或脂质组学研究。