Shanghai Jiaotong University, Shanghai, PR China.
J Microbiol Methods. 2012 May;89(2):102-6. doi: 10.1016/j.mimet.2012.02.009. Epub 2012 Feb 21.
Bartonella infection (Bartonella henselae in particular) is responsible for a widening spectrum of human diseases. The persistent colonization of erythrocytes is a feature of Bartonella infection. Endothelial and epithelial cells are also widely used to study the pathogenesis of bartonellosis in vitro. Exploring a convenient method for visualizing the bacillus without affecting infectivity would be very interesting. Carboxyfluorescein diacetate succinimidyl ester (CFSE) has been previously used for staining several bacterial species to study their adhesion to host cells. The present study demonstrated the efficiency and safety of using CFSE in staining B. henselae. The staining of bacillus-invaded erythrocytes and epithelial cells in vitro successfully allowed for flow cytometry and confocol microscopy analyses. Parallel tests using untreated bacteria confirmed that CFSE staining did not result in side effects on the infectivity of B. henselae. Labeling Bartonella with CFSE is a valuable method for studying the bacteria-host interaction.
巴尔通体感染(尤其是汉塞巴尔通体)可引起广泛的人类疾病。红细胞的持续定植是巴尔通体感染的一个特征。内皮细胞和上皮细胞也被广泛用于体外研究巴尔通体病的发病机制。探索一种不影响感染力的方便方法来可视化杆菌将非常有趣。羧基荧光素二乙酸琥珀酰亚胺酯(CFSE)以前曾用于对几种细菌进行染色,以研究它们对宿主细胞的粘附作用。本研究证明了使用 CFSE 对亨氏巴尔通体进行染色的效率和安全性。体外染色被杆菌入侵的红细胞和上皮细胞成功地允许进行流式细胞术和共聚焦显微镜分析。使用未经处理的细菌进行的平行测试证实,CFSE 染色不会对汉塞巴尔通体的感染力产生副作用。用 CFSE 标记巴尔通体是研究细菌-宿主相互作用的一种有价值的方法。