Institut Pasteur, BCI, Paris 75015, France.
Microbiol Spectr. 2019 Mar;7(2). doi: 10.1128/microbiolspec.BAI-0017-2019.
The spatial dimensions of host cells and bacterial microbes are perfectly suited to being studied by microscopy techniques. Therefore, cellular imaging has been instrumental in uncovering many paradigms of the intracellular lifestyle of microbes. Initially, microscopy was used as a qualitative, descriptive tool. However, with the onset of specific markers and the power of computer-assisted image analysis, imaging can now be used to gather quantitative data on biological processes. This makes imaging a driving force for the study of cellular phenomena. One particular imaging modality stands out, which is based on the physical principles of fluorescence. Fluorescence is highly specific and therefore can be exploited to label biomolecules of choice. It is also very sensitive, making it possible to follow individual molecules with this approach. Also, microscopy hardware has played an important role in putting microscopy in the spotlight for host-pathogen investigations. For example, microscopes have been automated for microscopy-based screenings. A new generation of microscopes and molecular probes are being used to image events below the resolution limit of light. Finally, workflows are being developed to link light microscopy with electron microscopy methods via correlative light electron microscopy. We are witnessing a golden age of cellular imaging in cellular microbiology.
宿主细胞和细菌微生物的空间维度非常适合通过显微镜技术进行研究。因此,细胞成像在揭示微生物细胞内生活方式的许多范例方面发挥了重要作用。最初,显微镜被用作定性、描述性工具。然而,随着特异性标记物的出现和计算机辅助图像分析的强大功能,成像现在可以用于收集生物过程的定量数据。这使得成像成为研究细胞现象的驱动力。一种特殊的成像模式脱颖而出,它基于荧光的物理原理。荧光具有高度特异性,因此可以用来标记选择的生物分子。它也非常灵敏,使得可以通过这种方法跟踪单个分子。此外,显微镜硬件在将显微镜置于宿主-病原体研究的聚光灯下方面发挥了重要作用。例如,显微镜已经自动化,用于基于显微镜的筛选。新一代显微镜和分子探针被用于对低于光分辨率的事件进行成像。最后,正在开发工作流程,通过相关的光电子显微镜将明场显微镜与电子显微镜方法联系起来。我们正见证细胞微生物学中细胞成像的黄金时代。