Kolek Jan, Branska Barbora, Drahokoupil Marek, Patakova Petra, Melzoch Karel
Department of Biotechnology, University of Chemistry and Technology Prague, Technicka 5, 16628 Prague, Czech Republic
Department of Biotechnology, University of Chemistry and Technology Prague, Technicka 5, 16628 Prague, Czech Republic.
FEMS Microbiol Lett. 2016 Mar;363(6). doi: 10.1093/femsle/fnw031. Epub 2016 Feb 8.
Flow cytometry, in combination with fluorescent staining, was used to evaluate population heterogeneity in acetone-butanol-ethanol fermentation that was carried out with type strain Clostridium beijerinckii NCIMB 8052 and non-type C. pasteurianum NRRL B-598. A combination of propidium iodide (PI) and carboxyfluorescein diacetate (CFDA), PI plus Syto-9 and bis-oxonol (BOX) alone were employed to distinguish between active and damaged cells together with simultaneous detection of spores. These strategies provided valuable information on the physiological state of clostridia. CFDA and PI staining gave the best separation of four distinct subpopulations of enzymatically active cells, doubly stained cells, damaged cells and spores. Proportional representation of cells in particular sub-regions correlated with growth characteristics, fermentation parameters such as substrate consumption and product formation in both species under different cultivation conditions.
流式细胞术结合荧光染色，用于评估丙酮-丁醇-乙醇发酵过程中的群体异质性，该发酵过程使用模式菌株拜氏梭菌NCIMB 8052和非模式菌株巴斯德梭菌NRRL B-598进行。碘化丙啶（PI）和羧基荧光素二乙酸酯（CFDA）的组合、单独的PI加Syto-9和双苯氧翁（BOX）用于区分活细胞和受损细胞，同时检测孢子。这些策略提供了有关梭菌生理状态的有价值信息。CFDA和PI染色能最好地分离出四个不同的亚群，即酶活性细胞、双染细胞、受损细胞和孢子。在不同培养条件下，特定亚区域细胞的比例与两种菌株的生长特征、发酵参数（如底物消耗和产物形成）相关。
