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一种用于实时追踪侵袭性细菌的简单高效荧光标记方法。

A simple and efficient fluorescent labeling method in for real-time tracking of invasive bacteria.

作者信息

Liu Fei, Chen Sijie, Zou Yingxin, Jiao Yong, Tang Ying

机构信息

Naval Medical Center, Naval Medical University, Shanghai, China.

Department of Nursing, The 940th Hospital of Joint Logistic Support Force of People's Liberation Army (PLA), Lanzhou, China.

出版信息

Front Microbiol. 2023 Feb 10;14:1128638. doi: 10.3389/fmicb.2023.1128638. eCollection 2023.

Abstract

Bacterial fluorescent labeling is a powerful tool for the diagnosis and treatment of bacterial infections. Here, we present a simple and efficient labeling strategy for . Intracellular labeling of bacteria was achieved by heat shock using Cyanine 5.5 (Cy5.5) near-infrared-I dyes in (). Several key factors, such as Cy5.5 concentration and labeling time, were systematically evaluated. Further, the cytotoxicity of Cy5.5 and the stability of was evaluated by flow cytometry, inverted fluorescence microscopy, and transmission electron microscopy. In addition, were used to explore the phagocytic behavior of RAW264.7 macrophages. These results proved that had a uniform fluorescence intensity and high luminance; additionally, our method had no significant adverse effects on compared to unlabeled infections. Our method provides researchers with a useful option for analyzing the behavior of as an infectious agent. This technique can be broadly applied to study host cell-bacteria interactions at the molecular level, and to tracing of bacterial infections.

摘要

细菌荧光标记是用于细菌感染诊断和治疗的有力工具。在此,我们提出一种简单有效的标记策略用于……通过在……中使用近红外-I染料花菁5.5(Cy5.5)进行热休克实现细菌的细胞内标记。系统评估了几个关键因素,如Cy5.5浓度和标记时间。此外,通过流式细胞术、倒置荧光显微镜和透射电子显微镜评估了Cy5.5的细胞毒性和……的稳定性。另外,……用于探究RAW264.7巨噬细胞的吞噬行为。这些结果证明……具有均匀的荧光强度和高亮度;此外,与未标记的……感染相比,我们的方法对……没有显著的不良影响。我们的方法为研究人员分析……作为感染因子的行为提供了一个有用的选择。该技术可广泛应用于在分子水平研究宿主细胞-细菌相互作用,以及用于……细菌感染的追踪。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c9c/9950555/ee4e7ed89569/fmicb-14-1128638-g001.jpg

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