微生物扫描电镜样品清洗方法的改进与固定干燥方法比较.pdf

安徽农业科学 , Journal of Anhu i A gri. Sci. 2009 , 37 (23) : 10886 - 10888 责任编辑 　庆瑢 　责任校对 　卢瑶 微生物扫描电镜样品清洗方法的改进与固定干燥方法比较 钱天乐 ,周逸卿 ,邹珍友 ,王 琰 ,孔肖菡 ,史永红 ,茅冬梅 ,谭同和 ,丁海东 ,郭 凯 ,赵 静 　 (南京农业大学生命科学实验中心 - 天美电镜技术有限公司合作示范实验室 ,江苏南京 2 10095) 摘要　以大肠杆菌为代表材料 ,用扫描电镜观察比较用 PB S和蒸馏水清洗再制样的图象效果 ,对比固定与不固定 ,用自然晾干法、烘干 法和用梯度酒精脱水再冷冻干燥法处理的大肠杆菌图像 。提出用蒸馏水替代 PB S清洗微生物样品以避免干燥后被固定液和 PB S残留 物覆盖的方法 , 明确具有细胞壁的细菌也需固定 ,对单细胞生物用梯度酒精脱水再冷冻干燥要比自然晾干和烘干脱水更彻底 。 关键词　扫描电镜;样品制作 ;微生物 ;大肠杆菌 ( ) 中图分类号　O657. 3; Q336　　文献标识码　A　　文章编号　0517 - 6611 2009 23 - 10886 - 03 Im provem en t of Ba th ing M ethod of M icroorgan ism Sam ples for SEM and Com par ison of The ir F ix ing and D ryness M ethods ( Q IA N T ian le et a l　 Coop erative Laboratory of Exp erim ental Center of L ife Science College and Techcom Corp. , N anj ing A gricu ltural Un i versity, N anj ing, J iangsu 2 10095) A b stract　W ith Escherich ia coli a s rep re sentative m aterials, the im age effects of Escherich ia coli after being bathed w ith BPS and distilledwa ter were ob served by u sing SEM. The conform ation s of the fixed and nonfixed samp le s were exam ined. The aired, warm incubated and freez inglyevaporated bacilli were comp ared. The re su lts indicated that water bathed samp le wa s more clear than that w ith PB S, for water sweep ed away the re sidue s of fixing so lu tion and PB S wh ich left on and covered the surface of samp les when dried. A lthough the cell wall can m aintain the conform ation , the bacterium can secrete out its inner sub stance s if do not fix. D ehydrating w ith the serie s of concentrate increa sing achol and then freezingly evaporating were better ways than airing or warm incubating for dryne s. Key words　Scann ing electric m icro scopy; Samp le p rep aring; M icroorgan ism ; Escherich ia coli 　　用常规方