4种观察副溶血弧菌表面形态的方法比较

Comparison between Four Methods for the Observation of Morphology of Vibrio Parahaemolyticus

以副溶血弧菌为材料,进行了相差显微镜、荧光显微镜、扫描电子显微镜、透射电子显微镜(负染)的观察,并得到清晰的菌体外表面形态的图片,同时分析比较各种方法的适用范围和优缺点,并对操作过程中可能遇到的问题以及需要注意的事项分别进行了阐述和说明。相差显微镜可以用于观察菌体的运动状况和单个菌体的大体形态,但无法观察细胞的微小变化;荧光显微镜可用于细菌计数、区分正常细胞和死细胞以及菌体细胞之间的连接现象和外分泌物的宏观情况等;扫描电子显微镜可观察到菌体细胞表面的微观形态,包括细胞壁褶皱等结构的细微变化,但不适于鞭毛和外分泌物的观察;透射电子显微镜(负染)可观察细菌活体的形态,可以对鞭毛和外分泌物等进行观察,但是不适用于需要表型统计的实验。

Vibrio parahaemolyticus was taken as the material to conduct observation by phase-contrast microscopy,fluorescence microscopy,scanning electron microscopy and transmission electron microscopy（negative staining） separately,and the clear pictures of outer surface of bacteria were obtained to analyze the sphere of application of these four methods and compare the advantages and disadvantages of them.Meanwhile,the possible encountered problems during operation and the key points that need to pay attention to were described separately.Phase-contrast microscope can be used to observe the movement of bacteria and the general shape of single bacteria,but can not observe the small changes of cell;fluorescence microscope can be used for counting,distinguishing between normal cells and dead cells,observing the phenomenon of connection between bacteria cells and external secretions and so on;scanning electron microscope can be used to observe the micro-morphology of bacterial cell surface,including the subtle fold changes of the cell wall structure,but not suitable for observation of flagella and the outer secretion;transmission electron microscope（negative staining） could used to observe the morphology of living bacteria,the flagella and the outer secretion,but not suitable for the experiment that require table-based statistics.