Electrospun polycaprolactone nanofibers were selectively ablated to form microstructures via the phase modulation of femtosecond laser beams. Ablation width (1-15 mu m) and depth (15-110 mu m) resolution were dependent upon the selection of pulse energy and microscope objective. Because phase modulation shapes light in a maskless fashion, desired templates were digitally created and physically transferred to electrospun mats within a matter of minutes. Several microarchitectures were formed in parallel by dividing pulse energy between multiple foci, substantially increasing throughput. The data presented herein demonstrates that phase-based laser ablation can be used to rapidly shape and tailor electrospun mats in three dimensions. (C) 2011 Elsevier B.V. All rights reserved.