Lippé Roger
Department of Pathology and Cell Biology, University of Montreal, Montreal, Québec, Canada
J Virol. 2018 Jan 17;92(3). doi: 10.1128/JVI.01765-17. Print 2018 Feb 1.
For several decades, flow cytometry has been a common approach to analyze cells and sort them to near-purity. It enables one to probe inner cellular molecules, surface receptors, or infected cells. However, the analysis of smaller entities such as viruses and exocytic vesicles has been more difficult but is becoming mainstream. This has in part been due to the development of new instrumentation with resolutions below that of conventional cytometers. It is also attributed to the several means employed to fluorescently label viruses, hence enabling them to stand out from similarly sized particles representing background noise. Thus far, more than a dozen different viruses ranging in size from 40 nm to giant viruses have been probed by this approach, which was recently dubbed "flow virometry." These studies have collectively highlighted the breadth of the applications of this method, which, for example, has elucidated the maturation of dengue virus, served as quality control for vaccinia vaccines, and enabled the sorting of herpes simplex virus discrete viral particles. The present review focuses on the means employed to characterize and sort viruses by this powerful technology and on the emerging uses of flow virometry. It similarly addresses some of its current challenges and limitations.
几十年来,流式细胞术一直是分析细胞并将其分选至接近纯态的常用方法。它能让人们探测细胞内部分子、表面受体或受感染细胞。然而,对病毒和胞外囊泡等较小实体的分析则更具难度,但正逐渐成为主流。这在一定程度上得益于分辨率低于传统细胞仪的新仪器的开发。这也归因于用于对病毒进行荧光标记的多种方法,从而使它们能够从代表背景噪声的大小相似的颗粒中凸显出来。到目前为止,这种方法已探测了十几种大小从40纳米到巨型病毒不等的不同病毒,该方法最近被称为“流式病毒分析”。这些研究共同突显了该方法应用的广度,例如,它阐明了登革病毒的成熟过程,用作痘苗疫苗的质量控制,并实现了单纯疱疹病毒离散病毒颗粒的分选。本综述重点关注通过这种强大技术对病毒进行表征和分选所采用的方法以及流式病毒分析的新用途。它同样探讨了其当前面临的一些挑战和局限性。