Department of Environmental Microbiology, Helmholtz Centre for Environmental Research-UFZ, Permoserstr. 15, 04318 Leipzig, Germany.
Cytometry A. 2010 Jul;77(7):623-34. doi: 10.1002/cyto.a.20920.
Single cell techniques like flow cytometry combined with viability staining can help to obtain information on viability states of bacteria. Many fluorescent dyes are available for this purpose and can be chosen according to the available excitation source, the species used, and the background of scientific questions and relevant specifications. Within this short overview, we focus on two diverse groups of bacteria: the gram- Escherichia coli and representatives of the gram+ Mycobacterium to demonstrate differences and similarities in dye uptake principles, processing and binding. We call for attention to possible diverse responses of different species to various viability assays. The cell surface structure of bacteria and the chemical properties of fluorescent probes considerably determine the success of a certain staining practice. Particular focus was drawn on analysis of membrane integrity, uptake of substrates and transformation of fluorogenic substrates.
单细胞技术,如流式细胞术结合活细胞染色,可以帮助获取细菌活力状态的信息。有许多荧光染料可用于此目的,可以根据可用的激发源、使用的物种、科学问题的背景和相关规范来选择。在这个简短的概述中,我们专注于两种不同的细菌群体:革兰氏阴性的大肠杆菌和革兰氏阳性的分枝杆菌,以展示在染料摄取原理、处理和结合方面的差异和相似性。我们呼吁注意不同物种对不同活力检测方法可能存在的不同反应。细菌的细胞表面结构和荧光探针的化学性质极大地决定了某种染色实践的成功。特别关注的是细胞膜完整性分析、底物摄取和荧光底物转化。