流式细胞术
碘杂醇
生物
色谱法
分析超速离心
细菌
超离心机
革兰氏阳性菌
小泡
密度梯度
微生物学
膜
生物化学
分子生物学
化学
量子力学
医学
物理
放射科
造影剂
遗传学
作者
Tadahiro Nasukawa,Ryosuke Sugimoto,Jumpei Uchiyama,Iyo Takemura‐Uchiyama,Hironobu Murakami,Ken Fukuda,Shigenobu Matsuzaki,Masahiro Sakaguchi
标识
DOI:10.1016/j.resmic.2020.11.003
摘要
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.
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