This review discusses the exploration of chromosome structure with a recently developed high-resolution microscopy technique, three-dimensional structured illumination microscopy (3dSIM). 3dSIM surpasses the diffraction limit of conventional widefield optical microscopy, increasing the level of detail in images by a factor of 2, while retaining the sample preparation methods, ease of use and flexibility of conventional microscopy. Special attention will be given to the ways in which imaging beyond the diffraction limit can shed light on the structural organization of meiotic chromosomes.