【摘要】 目的实时观察铜绿假单胞菌生物膜形成并测量其厚度。方法将绿色荧光蛋白表达质粒转化铜绿假单胞菌,再采用激光共聚焦显微镜对活的生物膜进行水平逐层扫描,观察生物膜形成过程与形态,根据三维坐标系Z轴距离计算生物膜厚度。结果铜绿假单胞菌生物膜形成过程分3个阶段:0~6 h为黏附阶段;6~24 h为聚集阶段,24~72 h为成熟阶段;6,24,48和72 h的生物膜厚度分别为(6.1±2.8),(29.2±2.3),(61.4±1.4),(61.8±1.1)μm。统计学分析表明,生物膜厚度6 h<24 h<48 h,差异有统计学意义(均P<0.01),48与72 h的生物膜厚度差异无统计学意义(P>0.05)。结论激光共聚焦实时观察法能够用于铜绿假单胞菌生物膜形成检测以及生物膜厚度测量。更多还原

【Abstract】 Objective To conduct real-time observation of living biofilm formation and determine the thickness of the Pseudomonas aeruginosa biofilm.Methods After green fluorescent protein(GFP)expression plasmid was transfected to Pseudomonas aeruginosa,the horizontal section scanning of living biofilm was done in real-time with confocal laser scanning microscope(CLSM).The thickness of living biofilm was determined according to Z axial distance.Results The biofilm formation of Pseudomonas aeruginosa was observed in three stages,which were the adherence phase from 0 to 6 hours,the assembly phase from 6 to 24 hours,and the maturation phase from 24 to 72 hours.The thickness of the biofilm at 6,24,48,and 72 hr were 6.1±2.8,29.2±2.3,61.4±1.4,and 61.8±1.1μm,respectively.Statistical analysis showed that the thickness of the biofilm at 6 hr was less than that of at 24 hr(P<0.01)and the thickness of the biofilm at 24 hr was less than that of at 48 hr(P<0.01).There was no difference in the thickness of the biofilm between the time of 48 hr and 72 hr(P>0.05).Conclusion CLSM can be used to observe the biofilm formation of Pseudomonas aeruginosa in real-time and to determine its thickness.更多还原