Carboxyfluorescein diacetate succinimidyl ester labeling method to study the interaction between Leptospira and macrophages
Carboxyfluorescein diacetate succinimidyl ester labeling method to study the interaction between Leptospira and macrophages
复制标题
羧基荧光素二乙酸琥珀酰亚胺酯标记法研究钩端螺旋体与巨噬细胞的相互作用。
DOI：
10.1016/j.mimet.2014.10.005
复制
发表时间：
2014-12-01
影响因子：
2.2
通讯作者：
He, Ping
中科院分区：
文献类型：
--
作者：
Liu, Boyu;Wang, Yanchun;He, Ping
钩端螺旋体病是由钩端螺旋体属致病性物种引起的，已成为世界上最广泛的人畜共患病之一。其确切的发病机制尚不清楚，钩端螺旋体与巨噬细胞之间的相互作用也不清楚。在这项研究中，我们报告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定量CFDA-SE标记的与巨噬细胞相关的钩端螺旋体。此外，我们发现台盼蓝有效地淬灭了附着的钩端螺旋体的细胞外荧光，这使得细胞内和细胞外细菌被区分开。(C)2014爱思唯尔有限公司版权所有。
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. (C) 2014 Elsevier B.V. All rights reserved.