Sandt C, Smith-Palmer T, Pink J, Brennan L, Pink D
Department of Chemistry, St Francis Xavier University, Antigonish, NS, Canada.
J Appl Microbiol. 2007 Nov;103(5):1808-20. doi: 10.1111/j.1365-2672.2007.03413.x.
To investigate the use of confocal Raman microspectroscopy (CRM) for the analysis of the structure, composition and development of fully hydrated biofilms.
Pseudomonas aeruginosa PAO1 biofilms were cultured in a flow cell in minimal nutrient medium (artificial sea water) and their development was followed for up to 3 weeks. The spectroscopic signature of the biofilm cells and extracellular polymeric substances (EPS) were differentiated and their distribution in biofilm colonies and within water channels was mapped in-plane and -depth. The colonies were initially amorphous, mainly composed of cells with no detectable amount of EPS. They developed rapidly to give round colonies composed of a cellular core enclosed in a sheath of EPS. The EPS continued to increase and spread throughout the biofilm to become the dominating feature of aged colonies. Colonies with a liquid core morphology - characteristic of the seeding dispersal process - were also observed.
This study demonstrated that CRM can be used to monitor the distribution of biofilm components in fully hydrated undisturbed biofilms over time.
Confocal Raman microspectroscopy facilitates the analysis of hydrated, live bacterial biofilms as a function of space and time, thus making it a suitable technique for investigating the effects of various additives and environmental factors on biofilm growth.
研究共聚焦拉曼显微光谱（CRM）在分析完全水合生物膜的结构、组成和发育方面的应用。
铜绿假单胞菌PAO1生物膜在流动池中于基本营养培养基（人工海水）中培养，并跟踪其长达3周的发育过程。区分了生物膜细胞和胞外聚合物（EPS）的光谱特征，并绘制了它们在生物膜菌落和水通道内的平面和深度分布。菌落最初是无定形的，主要由未检测到EPS的细胞组成。它们迅速发育成由包裹在EPS鞘中的细胞核心组成的圆形菌落。EPS持续增加并扩散到整个生物膜中，成为老化菌落的主要特征。还观察到具有液芯形态的菌落——这是种子传播过程的特征。
本研究表明，CRM可用于随时间监测完全水合且未受干扰的生物膜中生物膜成分的分布。
共聚焦拉曼显微光谱有助于分析水合的活细菌生物膜随空间和时间的变化，因此使其成为研究各种添加剂和环境因素对生物膜生长影响的合适技术。
