Actin restructuring during Salmonella typhimurium infection investigated by confocal and super-resolution microscopy
Actin restructuring during Salmonella typhimurium infection investigated by confocal and super-resolution microscopy
复制标题
DOI：
10.1117/1.jbo.19.1.016011
复制
发表时间：
2014-01-01
影响因子：
3.5
通讯作者：
Werner，James H.
中科院分区：
文献类型：
--
作者：
Han，Jason J.;Kunde，Yuliya A.;Werner，James H.
我们使用超分辨率光学显微镜和共聚焦显微镜来观察伴随并使鼠伤寒沙门氏菌内化的HeLa细胞的细胞骨架重组。在此，我们报告使用共聚焦显微镜来验证和探索与超分辨率光学显微镜兼容的感染条件，使用Alexa-488标记的phalloidin染色肌动蛋白细胞骨架网络。虽然众所周知，肌动蛋白重组和细胞骨架重排经常伴随和辅助细菌感染，但大多数研究都使用传统的衍射限制荧光显微镜来探索这些变化。本研究表明，单分子定位方法（如直接随机光学重建显微镜）提供的优越空间分辨率能够更精确地显示伴随细菌感染的肌动蛋白细胞骨架的纳米级变化。特别是，我们发现在感染后10到20分钟，一个薄的（100纳米）肌动蛋白环通常包围着入侵细菌，这个环在本质上是短暂的。我们估计有几百条肌动蛋白单丝围绕着这些细胞。鼠伤寒嘧啶：迄今未报道的细菌内化中间体。
We have used super-resolution optical microscopy and confocal microscopy to visualize the cytoskeletal restructuring of HeLa cells that accompanies and enablesSalmonella typhimuriuminternalization. Herein, we report the use of confocal microscopy to verify and explore infection conditions that would be compatible with super-resolution optical microscopy, using Alexa-488 labeled phalloidin to stain the actin cytoskeletal network. While it is well known that actin restructuring and cytoskeletal rearrangements often accompany and assist in bacterial infection, most studies have employed conventional diffraction-limited fluorescence microscopy to explore these changes. Here we show that the superior spatial resolution provided by single-molecule localization methods (such as direct stochastic optical reconstruction microscopy) enables more precise visualization of the nanoscale changes in the actin cytoskeleton that accompany bacterial infection. In particular, we found that a thin (100-nm) ring of actin often surrounds an invading bacteria 10 to 20 min postinfection, with this ring being transitory in nature. We estimate that a few hundred monofilaments of actin surround theS. typhimuriumin this heretofore unreported bacterial internalization intermediate.