National Centre for Advanced Tribology at Southampton (nCATS), Engineering Sciences, University of Southampton, Southampton SO17 1BJ, United Kingdom.
Bioengineering Sciences Group, Engineering Sciences, University of Southampton, Southampton SO17 1BJ, United Kingdom.
Biomicrofluidics. 2013 Dec 23;7(6):64118. doi: 10.1063/1.4850796.
In the current study, we have developed and fabricated a novel lab-on-a-chip device for the investigation of biofilm responses, such as attachment kinetics and initial biofilm formation, to different hydrodynamic conditions. The microfluidic flow channels are designed using computational fluid dynamic simulations so as to have a pre-defined, homogeneous wall shear stress in the channels, ranging from 0.03 to 4.30 Pa, which are relevant to in-service conditions on a ship hull, as well as other man-made marine platforms. Temporal variations of biofilm formation in the microfluidic device were assessed using time-lapse microscopy, nucleic acid staining, and confocal laser scanning microscopy (CLSM). Differences in attachment kinetics were observed with increasing shear stress, i.e., with increasing shear stress there appeared to be a delay in bacterial attachment, i.e., at 55, 120, 150, and 155 min for 0.03, 0.60, 2.15, and 4.30 Pa, respectively. CLSM confirmed marked variations in colony architecture, i.e.,: (i) lower shear stresses resulted in biofilms with distinctive morphologies mainly characterised by mushroom-like structures, interstitial channels, and internal voids, and (ii) for the higher shear stresses compact clusters with large interspaces between them were formed. The key advantage of the developed microfluidic device is the combination of three architectural features in one device, i.e., an open-system design, channel replication, and multiple fully developed shear stresses.
在当前的研究中,我们开发并制造了一种新型的片上实验室设备,用于研究生物膜对不同流体动力学条件的响应,如附着动力学和初始生物膜形成。微流道采用计算流体动力学模拟设计,以在通道中产生预定的、均匀的壁面剪切应力,范围为 0.03 至 4.30 Pa,这与船体以及其他人造海洋平台上的服役条件相关。使用延时显微镜、核酸染色和共聚焦激光扫描显微镜(CLSM)评估微流控装置中生物膜形成的时间变化。随着剪切应力的增加,观察到附着动力学的差异,即随着剪切应力的增加,细菌附着似乎会延迟,即在 0.03、0.60、2.15 和 4.30 Pa 时,分别为 55、120、150 和 155 分钟。CLSM 证实了菌落结构的明显变化,即:(i)较低的剪切应力导致生物膜具有独特的形态,主要特征为蘑菇状结构、间质通道和内部空隙;(ii)对于较高的剪切应力,形成了具有大间隔的紧密集群。所开发的微流控装置的主要优势在于在一个装置中结合了三个结构特征,即开放式设计、通道复制和多个完全发展的剪切应力。