摘要：
Fluorescence microscopy is a widely used imaging method in biological research due to its noninvasive nature and molecular specificity. The resolution of conventional fluorescence microscopy is limited to a few hundred nanometers by the diffraction of light, leaving many biological structures too small to be optically resolved. Stochastic Optical Reconstruction Microscopy (STORM) technique overcomes this limit by localizing single photoswitchable fluorophores separated in time. We further developed three-dimensional (3D) STORM by incorporating single molecule localization in 3D using astigmatism imaging, and demonstrated a 50-60 nm axial and 20-30 nm lateral resolution in mammalian cells. Enabled by the super-resolution imaging capability, we used 3D STORM along with biochemical assays to study the chromosome organization and cell wall ynthesis machinery in live bacteria.