This article presents an overview focusing on the structural control of electrospun membranes on a multilevel scale ranging from the morphology of single nanofibers to the packing and alignment of nanofibers and the patterns and shapes of fibrous scaffolds. The typical structures of electrospun membranes and the specific electrospinning strategies used to produce these structures are reviewed. In addition, potential applications of these controlled structures in tissue engineering and drug delivery are highlighted. Finally, this review concludes with a perspective on the challenges and future directions for the design and fabrication of electrospun scaffolds using controlled structures along with an investigation of the relationship between the structures of electrospun membranes and the cell and drug delivery behaviors.