Single-molecule fluorescence imaging visualizes biomolecules at high spatial and temporal resolution, but it is limitedly used for studying cell surfaces or small areas inside cells. Jialei Tang, a graduate student in Optical Nanoscopy lab led by Dr. Kyu Young Han overcame the current limitation of single-molecule imaging approaches, and demonstrated clear and large field-of-view 3D single-molecule microscopy capable of imaging multiple cells and brain tissues. This will benefit several applications including super-resolution imaging, high-throughput gene expression profiling and single-molecule tracking. This work was published in Optica.