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Flow cytometry and FACS applied to filamentous fungi

机译：流式细胞术和FACS适用于丝状真菌

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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 (FAGS) 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 'omits' 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. (C) 2018 Published by Elsevier Ltd on behalf of British Mycological Society.
机译：流式细胞术是一种自动化的激光或阻抗的高通量方法，其允许在细胞群内的单细胞的多种化学和物理特性进行非常快速的分析。这是一种极其强大的技术，已被使用超过四十年的丝状真菌。尽管使用该技术通常在细胞逐细胞内快速分析细胞群内的单细胞，但流式细胞术也可用于分析细胞（例如孢子）聚集体或整个微菌根。可以用各种荧光记者染色生物或固定细胞，以标记不同的细胞成分或测量不同的生理过程。流式细胞术也适用于通过分析光散射特性使用未标记的细胞测量细胞尺寸，相互作用，聚集或形状。荧光激活的细胞分选（FAG）是一种专用形式的流式细胞术，其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成两种或更多种容器的方法。通过流式细胞术分析细胞在显微镜下分析细胞的主要优点是分析速度：每秒可以分析数千个细胞或以分钟分选。流式细胞术的缺点是不能跟踪特定细胞，并且缺乏与单个细胞有关的空间信息。微观显微镜的一个大的优点是使用FACS时，可以对下游实验进行分类具有所需特性的细胞（例如，用于生长，感染，酶产生，基因表达测定或“省略”方法）。在本文中，我们解释了流式细胞术和FACS的基本概念，与显微镜相比，定义其优点和缺点，并描述了这些强大技术已被丝状真菌使用的广泛应用。 （c）2018年由elsevier有限公司发布代表英国Mycological社会。

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