Liu Boyu, Wang Yanchun, Guo Xiaokui, Zhu Weinan, Zhang Yan, He Ping
Department of Immunology and Microbiology, Institute of Medical Science, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
J Microbiol Methods. 2014 Dec;107:205-13. doi: 10.1016/j.mimet.2014.10.005.
Leptospirosis, which is caused by pathogenic species of the genus Leptospira, has emerged as one of the most widespread zoonotic diseases in the world. The exact mechanism of pathogenesis remains unknown, and the interaction between Leptospira and macrophages is not well understood. In this study, we report that carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) can efficiently label different Leptospira interrogans strains without affecting bacterial motility, viability, or virulence. Following co-incubation, CFDA-SE-labeled leptospires associated with macrophages were quantified by flow cytometry or confocal microscopy. In addition, we showed that trypan blue efficiently quenched the extracellular fluorescence from the adherent leptospires, which enabled intracellular and extracellular bacteria to be distinguished.
钩端螺旋体病由钩端螺旋体属的致病菌种引起，已成为世界上传播最广泛的人畜共患病之一。发病的确切机制尚不清楚，钩端螺旋体与巨噬细胞之间的相互作用也未得到充分了解。在本研究中，我们报告了羧基荧光素二乙酸琥珀酰亚胺酯（CFDA-SE）可以有效地标记不同的问号钩端螺旋体菌株，而不影响细菌的运动性、活力或毒力。共孵育后，通过流式细胞术或共聚焦显微镜对与巨噬细胞相关的CFDA-SE标记的钩端螺旋体进行定量。此外，我们还表明，台盼蓝能有效淬灭粘附钩端螺旋体的细胞外荧光，从而区分细胞内和细胞外细菌。
