Flow cytometry is an automated, laser- or impedance-based, high throughput method that allows very rapid analysis of multiple chemical and physical characteristics of single cells within a cell population. It is an extremely powerful technology that has been used for over four decades with filamentous fungi. Although single cells within a cell population are normally analysed rapidly on a cell-by-cell basis using the technique, flow cytometry can also be used to analyse cell (e.g. spore) aggregates or entire microcolonies. Living or fixed cells can be stained with a wide range of fluorescent reporters to label different cell components or measure different physiological processes. Flow cytometry is also suited for measurements of cell size, interaction, aggregation or shape using non-labelled cells by means of analysing their light scattering characteristics. Fluorescence-activated cell sorting (FACS) is a specialized form of flow cytometry that provides a method for sorting a heterogeneous mixture of cells into two or more containers based upon the fluorescence and/or light scattering properties of each cell. The major advantage of analysing cells by flow cytometry over microscopy is the speed of analysis: thousands of cells can be analysed per second or sorted in minutes. Drawbacks of flow cytometry are that specific cells cannot be followed in time and normally spatial information relating to individual cells is lacking. A big advantage over microscopy is when using FACS, cells with desired characteristics can be sorted for downstream experimentation (e.g. for growth, infection, enzyme production, gene expression assays or ‘omics’ approaches). In this review, we explain the basic concepts of flow cytometry and FACS, define its advantages and disadvantages in comparison with microscopy, and describe the wide range of applications in which these powerful technologies have been used with filamentous fungi.

流式细胞仪是一种自动化的，基于激光或基于阻抗的高通量方法，可以非常快速地分析细胞群中单个细胞的多种化学和物理特征。这是一项非常强大的技术，已与丝状真菌一起使用了四十年以上。尽管通常使用该技术在每个细胞的基础上快速分析细胞群中的单个细胞，但是流式细胞仪也可以用于分析细胞（例如孢子）聚集体或整个微菌落。活细胞或固定细胞可以用多种荧光报告基因染色，以标记不同的细胞成分或测量不同的生理过程。流式细胞仪还适用于通过分析未标记细胞的光散射特性来测量其细胞大小，相互作用，聚集或形状。荧光激活细胞分选（FACS）是流式细胞术的一种特殊形式，它提供了一种基于每个细胞的荧光和/或光散射特性将细胞的异质混合物分装到两个或多个容器中的方法。通过流式细胞术而不是显微镜分析细胞的主要优势是分析速度：每秒可分析成千上万个细胞，或在数分钟内将其分类。流式细胞术的缺点是不能及时跟踪特定的细胞，通常缺少与单个细胞有关的空间信息。与显微镜相比，最大的优势是当使用FACS时，可以将具有所需特征的细胞进行分选以进行下游实验（例如用于生长，感染，酶产生，基因表达分析或“组学”方法）。在这篇综述中，我们解释了流式细胞术和流式细胞仪的基本概念，定义了其与显微镜相比的优缺点，并描述了这些功能强大的技术已与丝状真菌一起使用的广泛应用。