Case Study: Illuminating the Nanoscale World of Microbiology.

Super-resolution microscopy has emerged as a groundbreaking tool in microbiology, enabling the visualization of structures and molecules far below the diffraction limit of light. We provide an overview of the various techniques employed in super-resolution microscopy, including deterministic (scanning and structured illumination) and stochastic (single-molecule localization microscopy and fluctuation-based computation) methods, summarizing their respective advantages and limitations. Applications in microbiology are presented, including investigations of cellular processes, physiology, cell wall biology, and elucidation of bacterial toxin pathogenesis mechanisms at nanoscale resolution. New and emerging uses of super-resolution microscopy are also explored, focusing on addressing critical challenges in the field: pathogen-host interactions, antibiotic resistance, mode of action of bacterial toxins, and nanoscale detail of bacterial secretion systems and their translocated effectors. With the expanding accessibility of resources and increasing availability of open-source software to democratize data analysis, super-resolution microscopy is poised to revolutionize our understanding of the nanoscale world in microbiology, paving the way for new discoveries.