论文部分内容阅读
目的 建立变异链球菌生物膜模型 ,用激光共聚焦扫描显微镜 (CLSM)观察变异链球菌生物膜结构。方法 在盖玻片上分别形成 6、12、18、2 4、4 8、72h变异链球菌生物膜 ,将得到的各时段生物膜荧光染色后 ,用CLSM观察生物膜的断层扫描图像、生物膜厚度、每层红光绿光的面积 ,计算生物膜中细菌密度和活菌百分比 ,用软件处理扫描数据 ,得到生物膜的三维重建图像。结果 变异链球菌生物膜具有空间立体结构 ,形态多样 ,其中细菌密集 ,由死细菌和活细菌组成 ,还有丰富的基质和管道系统。 2 4h生物膜平均厚度最大 ,生物膜内层、中间层的细菌密度相对较大 ,而外层较低 ,72h内各时间段生物膜中活菌百分比由内往外逐渐增加。结论 变异链球菌生物膜有一定的厚度 ,具有三维立体空间结构 ,结构形态具有多样、不均质、开放的特点。
Objective To establish a Streptococcus mutans biofilm model and observe the biofilm structure of Streptococcus mutans by laser scanning confocal microscope (CLSM). Methods The biofilms of Streptococcus mutans at 6, 12, 18, 24, 48 and 72h were formed on the coverslips respectively. After the biofilms were stained by CLSM, the biofilms were observed by CLSM. The biofilm thickness , The area of each layer of red-green light, calculate the bacterial density in the biofilm and the percentage of viable bacteria, and process the scan data by software to obtain the three-dimensional reconstructed image of the biofilm. Results Streptococcus mutans biofilm with spatial three-dimensional structure, shape and diversity, including bacteria dense, composed of dead bacteria and live bacteria, as well as a rich matrix and piping system. The average thickness of the biofilm was the highest at 24 h. The bacterial density in the inner and the middle layer of the biofilm was relatively large while the outer layer was relatively low. The percentage of viable biofilm in each biofilm increased gradually from the inside to the outside within 72 hours. Conclusion Streptococcus mutans biofilm has a certain thickness, with three-dimensional spatial structure, the structure of the shape of a diverse, heterogeneous, open characteristics.