[1]陈波曼,余加林,刘官信,等.SYTO9/PI荧光探针标记的铜绿假单胞菌PAO1菌株生物被膜形成的动态观察[J].陆军军医大学学报(原第三军医大学学报),2008,30(05):390-392.
 CHEN Bo-man,YU Jia-lin,LIU Guan-xin,et al.Sequential development of biofilm spatial structure of Pseudomonas Aeruginosa tagged with SYTO9/PI[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(05):390-392.
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SYTO9/PI荧光探针标记的铜绿假单胞菌PAO1菌株生物被膜形成的动态观察(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第05期
页码:
390-392
栏目:
论著
出版日期:
2008-03-15

文章信息/Info

Title:
Sequential development of biofilm spatial structure of Pseudomonas Aeruginosa tagged with SYTO9/PI
作者:
陈波曼余加林刘官信胡琳燕李芳杨华
浙江大学医学院附属妇产科医院新生儿科; 重庆医科大学附属儿童医院新生儿科
Author(s):
CHEN Bo-man YU Jia-lin LIU Guan-xin HU Lin-yan LI Fang YANG Hua
Department of Neonatology, Women Hospital, School of Medicine, Zhejiang University; Department of Neonatology, Children’s Hospital, Chongqing Medical University
关键词:
生物被膜铜绿假单胞菌定量分析结构
Keywords:
biofilm Pseudomonas Aeruginosa quantitative analysis structure
分类号:
Q93-31; Q935; Q939.112
文献标志码:
A
摘要:
目的 探讨铜绿假单胞菌细菌生物被膜(biofilm,BF)形成发展过程中空间立体结构变化。 方法 SYTO9/PI荧光探针标记PAO1菌株,体外建立6h、1d、3d及6d时间组铜绿假单胞菌PAO1菌株BF模型,激光共聚焦显微镜(confocal laser scanning microscopy, CLSM)摄取BF形成发展各阶段不同层面的图片堆,经图像结构分析软件(image structure analyzer, ISA)分析获得PAO1菌株BF相关空间结构参数定量化数据。 结果 ①CLSM结合SYTO9/PI荧光探针标记,实现了对PAO1菌株BF动态形成过程的观察。②随着BF形成时间的延长,各层死菌比例逐渐增加,且死菌多分布于微菌落中心。③ISA软件定量化分析显示,随着BF发展,厚度显著增加,前3d增加程度显著大于后3 d;区域孔率显著下降趋势;平均扩散距离呈逐渐增加趋势;结构熵呈显著上升趋势。 结论 利用ISA程序结合SYTO9/PI荧光探针可以表征PAO1菌株BF的空间结构特征和BF各层细菌的生存状态。
Abstract:
Objective To investigate the sequential development of spatial structure of biofilm with Image Structure Analyzer (ISA) software so as to found a basis for further research on biological behavior of biofilm.  MethodsIn vitro biofilm model of Pseudomonas Aeruginosa (P. aeruginosa) PAO1 tagged with SYTO9/PI was established on glass slice, and its biofilm development was monitored at different time points (6 h, 1, 3, and 6 d). The fluorescence images stack of different layers in biofilm model were obtained by confocal laser scanning microscopy (CLSM) based on fluorophores from PAO1. Quantitative parameters describing biofilm spatial structure were acquired after the image information was calculated by ISA software.  Results ①The PAO1 biofilm process was investigated successfully by CLSM after genetically tagged with GFP. ②With the development of biofilm, the ratio of died bacteria was increased gradually in each layer and most of them were distributed in the core of microcolony. ③The quantitative data from ISA software showed that the thickness of biofilm was increased significantly during biofilm growth; meanwhile the areal porosity (AP) was decreased significantly, the average diffusion distance (ADD) indicated an increased tendency; and the textural entropy (TE) increased significantly.  Conclusion The ISA software combined with SYTO9/PI tagging provide useful information of bacterial biofilm spatial structure in PAO1 biofilm process. Quantifying bacterial biofilm structure permits correlating biofilm development with biofilm performance.

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更新日期/Last Update: 2008-04-21