Multicolor Super-resolution Fluorescence Imaging of Cells by Expansion Microscopy

This chapter presents a step-by-step protocol to resolve the organization of the nuclear pore complex (NPC) in HeLa-NUP107-GFP cells by eight-fold expansion microscopy (ExM). To enhance the efficiency of the procedure, we introduce an accelerated expansion protocol designed to allow rapid iteration and improve experimental throughput. This protocol involves both pre-expansion staining with wheat germ agglutinin (WGA) and post-digestion staining with nanobodies against GFP to visualize the NPC. The subsequent use of confocal microscopy enables multicolor super-resolution imaging with approximately 30 nm spatial resolution.