Liu Xiaohui, Pomorski Thomas Günther, Liesche Johannes
College of Life Science, Northwest A&F University, Yangling, China.
Biomass Energy Center for Arid Lands, Northwest A&F University, Yangling, China.
Bio Protoc. 2019 Aug 20;9(16):e3344. doi: 10.21769/BioProtoc.3344.
All bacteria, fungi and plant cells are surrounded by a cell wall. This complex network of polysaccharides and glycoproteins provides mechanical support, defines cell shape, controls cell growth and influences the exchange of substances between the cell and its surroundings. Despite its name, the cell wall is a flexible, dynamic structure. However, due to the lack of non-invasive methods to probe the structure, relatively little is known about the synthesis and dynamic remodeling of cell walls. Here, we describe a non-invasive method that quantifies a key physiological parameter of cell walls, the porosity, , the size of spaces between cell wall components. This method measures the porosity-dependent decrease of the plasma membrane-localized fluorescent dye FM4-64 in the presence of the extracellular quencher Trypan blue. This method is applied to bacteria, fungi and plant cell walls to detect dynamic changes of porosity in response to environmental cues.
所有细菌、真菌和植物细胞都被细胞壁所包围。这种由多糖和糖蛋白组成的复杂网络提供机械支撑，确定细胞形状，控制细胞生长，并影响细胞与其周围环境之间的物质交换。尽管名为细胞壁，但它是一种灵活、动态的结构。然而，由于缺乏探测其结构的非侵入性方法，人们对细胞壁的合成和动态重塑了解相对较少。在此，我们描述了一种非侵入性方法，该方法可量化细胞壁的一个关键生理参数——孔隙率，即细胞壁成分之间空间的大小。此方法通过细胞外淬灭剂台盼蓝检测质膜定位荧光染料FM4 - 64因孔隙率而导致的减少。该方法应用于细菌、真菌和植物细胞壁，以检测孔隙率响应环境线索的动态变化。
