大肠杆菌长时间观察的方法

细菌个体微小,在液体中运动性强,且有的细菌有鞭毛、菌毛等结构,增加其运动能力,因此对感兴趣的细菌长时间观察很有难度.本文采用激光扫描共聚焦显微镜,比较多聚赖氨酸或低熔点琼脂糖处理后,大肠杆菌长时间观察的效果.结果发现在多聚赖氨酸包被的培养皿上加入含1.5%低熔点琼脂糖的LB培养基、室温培养或者在多聚赖氨酸包被的培养皿上加入含6%低熔点琼脂糖的LB培养基37℃培养,大肠杆菌均可在聚焦面贴壁分裂生长.连续观察2h以上大肠杆菌没有漂离视野,实现了显微镜下清晰呈现大肠杆菌分裂生长过程的目的.

Method for long-time imaging of E. coli

Long-time imaging can be used to observe the cell cycle, track the expression of specific proteins. Due to the small size and strong mobility in liquid, it is difficult to follow up interested bacteria for a long time under a microscope. In this study, E. coli was treated with poly-lysine or low melting agarose, and observed by laser scanning confocal microscope. The results reveal that E. coli could grow adhered to polylysine-coated dish containing 1. 5% low melting agarose medium at room temperature or topolylysine-coated plate containing 6% low melting agarose medium at 37 ℃. E. coli didn' t drift away from the field of vision after observation of more than two hours, and its growth process can be clearly observed under laser scanning confocal microscope.