原子力显微镜在活体微生物实时动态观测中的应用

Application of Atomic Force Microscopy in the in-situ Observation of Vital Microorganism

本文运用原子力显微镜的实时成像技术,通过探索制样条件和调节仪器参数,对活体大肠杆菌进行原位实时动态观测。文中探讨研究了生理盐水、PBS磷酸缓冲液以及Tris-HCl缓冲液洗涤之后细菌的形态变化。结果表明,采用Tris-HCl缓冲液洗涤后的细菌仍然保持良好的状态,菌体饱满光滑,没有出现干瘪塌陷或是形成多细胞聚集体的现象,能够真实反映细菌的形态以及表面微结构。另外,调节原子力显微镜回馈信号的积分增益和力学参数,比较长时间实时动态观测下细菌形貌的差异,优化仪器参数设置。经过两个小时的连续观察,细菌依然保持良好的状态,菌体完整饱满,表面平整光滑,实现了活体微生物的原位实时动态观测。

In this study,the in-situ observation of vital microorganism was carried out with atomic force microscopy.The influences of sample preparation and instrumental parameters were investigated.Effect of different buffers in rinse process was evaluated.Results showed that the bacteria were plump and the bacterial surface kept smooth after rinsing with Tris-HCl buffer.In addition,the influences of integral gain and the force setpoint in the feedback control of atomic force microscope were also discussed.With the optimized parameters,the bacteria kept in good condition during 2 hour continuous in-situ observation.