At only 50 nm in diameter, plasmodesmata (PD) are below the limit of resolution of conventional light microscopy. Consequently, much of our current interpretation of the substructure of PD is derived from transmission electron microscopy. However, PD can be imaged with alternative techniques, including field emission scanning electron microscopy and 'super-resolution' imaging approaches such as 3D-structured illumination microscopy. This review considers the methods currently available for studying PD and focuses on the boundary between light- and electron-based imaging approaches.