Alexa-555 WGA和FM4-64在活细菌定位中的比较应用  

Comparison Application of Alexa-555 WGA and FM4-64 in the Localization of Viable Bacteria

作　　者：王净[1,2] 韩延平[2] 杨瑞馥[2] 

机构地区：[1]河北北方学院动物科技学院,宣化075131 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《基因组学与应用生物学》2017年第7期2892-2896,共5页Genomics and Applied Biology

基　　金：国家自然科学基金(31171248);国家重点基础研究发展计划(973)项目(2014CB744405);河北省科技计划项目[14236602D-2(2015)]共同资助

摘　　要：为了准确定位亚细胞形态,本试验比较应用Alexa-555 WGA和FM4-64两种染料,为研究者合理选择染料提供线索。以大肠杆菌(Escherichia coli)、鲍曼不动杆菌(Acinetobacter baumannii)、金黄色葡萄球菌(Staphylococcus aureus)和炭疽芽孢杆菌(Bacillus anthracis)为对象,使用Hoechst 33342标记拟核,FM4-64标记细胞膜,Alexa-555 WGA标记细胞壁,Perkin Elmer Ultra VIEW Vo X荧光共聚焦显微镜观察成像。FM4-64染料适合于革兰氏阴性菌细胞膜染色,Alexa-555 WGA染料对革兰氏阴性菌和革兰氏阳性菌细胞壁染色效果均较好。Alexa-555 WGA能用于不同类型细菌细胞壁的定位。本研究首次将Alexa-555 WGA染料应用于原核生物,可为细菌的亚细胞定位研究提供借鉴。In order to accurately locate the subcellular morphology, in this experiment, we compared two kinds of dyes, Alexa-555 WGA and FM4-64, to provide clues for the researchers on choosing dyes reasonably. Escherichia coli, Acinetobacter baumannii, Staphylococcus aureus and Bacillus anthracis were selected as the object, we used Hoechst 33342 to label the nucleus, FM4-64 to label cell membrane, Alexa-555 WGA to label cell wall, and PerkinElmer Ultra VIEW VoX fluorescence confocal microscopy to observe the imaging. FM4-64 dyes was suit- able for gram negative bacterial cell membrane staining, and gram-negative and gram-positive bacteria were dyed better with Alexa-555 WGA dye. Alexa-555 WGA dye could be used to localize cell wall of different types of bacteria. Alexa-555 WGA dye was was applied to prokaryotes for the first time in this study, which could provide reference for studying subcellular localization.

关 键 词：Alexa-555 WGA FM4-64 活细菌 细胞壁 标记定位 

分 类 号：Q93[生物学—微生物学]