Assessment of change in biofilm architecture by nutrient concentration using a multichannel microdevice flow system
Assessment of change in biofilm architecture by nutrient concentration using a multichannel microdevice flow system
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DOI：
10.1016/j.jbiosc.2012.09.018
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发表时间：
2013-03-01
影响因子：
2.8
通讯作者：
Kimbara, Kazuhide
中科院分区：
文献类型：
--
作者：
Sanchez, Zoe;Tani, Akio;Kimbara, Kazuhide
采用一种新型的不锈钢流动室多通道微装置流动系统，对绿色荧光蛋白（GFP）标记的铜绿假单胞菌（Pseudomonasaeruginosa）PAO 1活生物膜进行了结构可视化、发育监测和结构定量。直接在原位调查，共聚焦激光扫描显微镜（CLSM）在72小时揭示了结构模式的差异，营养浓度梯度的结果。当生长在LB培养基中时，形成圆形的、分散的细胞聚集体，而在1/3稀释的LB培养基中，生物膜大多是平坦的和紧凑的。然而，COMSTAT分析表明，两个LB浓度之间的生物量和厚度没有相当大的差异。在1/3稀释的LB培养基中生长的生物膜的时间依赖性发育的表征显示，菌落在120小时时完全成熟，达到最大生物量17.1 μ m（3）/μ m（2），平均厚度为44.4 μ m。到168 h，菌落变薄，并形成穿过菌落内部的开口。这些结果表明，测试的新系统允许在不锈钢流动室的表面上快速且厚的生物膜发展。这些研究结果可以提供更好的估计生物膜活性和不同参数对生物膜形态和发展的影响，在工业和生物医学系统的系统评价。(C)2012年，生物技术学会，日本。All rights reserved.
A new multichannel microdevice flow system with stainless steel flow chamber was used for architecture visualization, development monitoring and structural quantification of GFP-labeled Pseudomonas aeruginosa PAO1 live biofilms. Direct in situ investigations using confocal laser scanning microscopy (CLSM) at 72 h revealed structural pattern differences as a result of nutrient concentration gradients. When grown in LB medium, round, dispersed cellular aggregates were formed whereas in 1/3-diluted LB medium, biofilms were mostly flat and compact. However, COMSTAT analyses showed no considerable differences in biomass and thickness between the two LB concentrations. Characterization of time-dependent development of biofilms grown in 1/3-diluted LB medium showed full maturation of colonies by 120 h reaching maximum biomass at 17.1 mu m(3)/mu m(2) and average thickness at 44.4 mu m. Consequent thinning and formation of openings through interior in colonies occurred by 168 h. These results suggest that the new system tested allowed a fast and thick biofilm development on the surface of the stainless steel flow chamber. These findings may provide better estimates of biofilm activity and systematic evaluation of the effects of different parameters on biofilm morphology and development in industrial and biomedical systems. (C) 2012, The Society for Biotechnology, Japan. All rights reserved.