Abstract：
The cariogenicity of Streptococcus mutans relates to its ability to form biofilms on dental surfaces. The aim of this work was to develop a flowcell system compatible with time-lapse confocal microscopy to compare the adhesion and accumulation of S. mutans cells on surfaces in unsupplemented media against media containing sucrose or sucralose (a non-metabolized sweetener) over a short period of time. Fluorescent S. mutans 3209/pVMCherry was suspended in unsupplemented media or media supplemented with 1% sucrose or 1% sucralose and passed through a 3D-printed flowcell system. Flowcells were imaged over 60 minutes using a confocal microscope. Image analysis was performed, including a newly developed object-movement-based method to measure biomass adhesion. Streptococcus mutans 3209/pVMCherry grown in 1% sucrose-supplemented media formed small, dense, relatively immobile clumps in the flowcell system measured by biovolume, surface area, and median object centroid movement. Sucralose-supplemented and un-supplemented media yielded large, loose, mobile aggregates. Architectural metrics and per-object movement were significantly different (P < 0.05) when comparing sucrose-supplemented media to either unsupplemented or sucralose-supplemented media. These results demonstrate the utility of a flowcell system compatible with time-lapse confocal microscopy and image analysis when studying initial biofilm formation and adhesion under different nutritional conditions.
摘要：
变形链球菌的致龋性与其在牙齿表面形成生物膜的能力有关。这项工作的目的是开发一种与延时共聚焦显微镜兼容的流通池系统，以比较变形链球菌细胞在未补充培养基中与含有蔗糖或三氯蔗糖（非代谢甜味剂）的培养基中的表面上的粘附和积累。短时间。将荧光变形链球菌3209/pVMCherry悬浮在未补充的培养基或补充有1%蔗糖或1%三氯蔗糖的培养基中,并通过3D打印的流通池系统。使用共聚焦显微镜在60分钟内对流式细胞成像。进行了图像分析，包括新开发的基于对象运动的方法来测量生物质粘附。在1%蔗糖补充培养基中生长的S.mutans3209/pVMCherry形成小的，密集,通过生物体积测量的流动池系统中相对固定的团块，表面积,和中间物体质心运动。三氯蔗糖补充和未补充的培养基产生了大量，松散,移动骨料。当比较补充蔗糖的培养基与未补充或补充三氯半乳蔗糖的培养基时,结构度量和每对象移动显著不同(p<0.05)。这些结果证明了在不同营养条件下研究初始生物膜形成和粘附时，与延时共聚焦显微镜和图像分析兼容的流动池系统的实用性。
