Developing super-resolution fluorescent microscopy for virology research.

Abstract

Taking advantage of the use of photoswitchable probes and high precision localisation of single molecules to surpass the diffraction limit, super-resolution fluorescence microscopy allows observing non-invasive live-cell at sub-diffraction size (<200 nm). Given the advantage of super-resolution fluorescence microscopy, our group has reconstructed the super-resolution fluorescence microscopy based on the single-molecule localisation microscopy technique with a resolution of 20 nm. In this research, the authors present the reconstruction process of the microscopy system and its application in observing hemorrhagic fever Dengue virus. Dengue virus was cultured in baby hamster kidney (BHK-21) cells and was then negative stained for transmission electron microscope (TEM) or immunofluorescent labeled for stochastic optical reconstruction microscopy (STORM). The diameter of the Dengue virus particles is 45-60 nm measured using TEM and is 84±12 nm measured using STORM. After subtraction of the length of the antibody attached to the virus particles, the diameter of Dengue virus particles measured using STORM are close to which measured using TEM. In conclusion, the authors highlight the findings of super-resolution fluorescence microscopy-based Dengue virus studies and their contributions to the understanding of Dengue virus particles. The current advances in super-resolution microscopy may open new avenues for future virology teaching and research.