超分辨率显微镜技术的比较,用于成像紧密包装的微殖民地,一个有义务的细胞内细菌
Alison J North1, Ved P Sharma1, Christina Pyrgaki1,2
1Bio-Imaging Resource Center, The Rockefeller University, New York, New York, USA.
Journal of microscopy
|December 9, 2024
概括
像3D-SIM和3D-STED这样的超高分辨率显微镜技术可以改善对Orientia tsutsugamushi (Ot) 细菌的成像. 将3D-STED与深度学习分析相结合,揭示了不同宿主细胞中的细菌形状变异.
科学领域:
- 显微镜和成像科学 显微镜和成像科学
- 细胞生物学 细胞生物学
- 细菌学 细菌学是一门学科.
背景情况:
- 由于细胞大小小小,凝聚和标记困难,对有义务的细胞内细菌进行成像是具有挑战性的.
- 传统的光学显微镜缺乏可视化像Orientia tsutsugamushi (Ot) 这样的细菌在宿主细胞内所需的分辨率.
研究的目的:
- 为了比较五种光显微镜技术的疗效,用于成像细胞内Orientia tsutsugamushi (Ot).
- 评估超分辨率方法,以分辨细菌细胞和外部囊泡在横向和轴向维度.
- 用先进的成像和分析分析不同哺乳动物细胞系内的细菌形状和大小变异.
主要方法:
- 使用共聚焦,Airyscan,iSIM,3D-SIM和STED显微镜对Orientia tsutsugamushi (Ot) 的成像.
- 使用ScaA蛋白的全宽半最大 (FWHM) 测量进行横向分辨率的定量评估.
- 应用3D-STED方法与Cellpose (深度学习) 和Imaris (商业软件) 结合进行细分和分析.
主要成果:
- 三维结构照明显微镜 (3D-SIM) 提供了最好的侧面分辨率,特别是使用短波长染料.
- 三维刺激辐射耗尽显微镜 (3D-STED) 提供了优越的轴分辨率.
- 基于深度学习的分析揭示了细菌形状的显著差异,但在各种宿主细胞系中没有大小差异.
结论:
- 超高分辨率显微镜,特别是3D-SIM和3D-STED显著提高了细胞内细菌的成像.
- 结合3D-STED和深度学习分析,可以有效地对宿主细胞内的细菌进行详细的形态学研究.
- 细菌的形状受到宿主细胞环境的影响,为宿主-病原体相互作用提供了新的见解.
关键词:
3D细胞细分3D细胞细分3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM 3D-SIM空中扫描可以扫描空中扫描.东方的 tsutsugamushi 东方的 tsutsugamushi 是一个瑞克特西亚尔斯 (Rickettsiales) 是一个小小的动物.子 一个子与焦点一致的对焦器深度学习是一种深度学习.这里是 iSIM.需要有义务的细胞内细菌超高分辨率成像成像技术
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