Candidate substrate designs for aligned electrospun PLLA nanofibers. Photos (A, C, E) and SEM images (B, D, F) of nanofibers spun onto glass cover slips. (A, B) Fibers were directly electrospun on cover slips attached to the rotating target wheel. No glue was used to fasten them to the cover slip. (C, D) Fibers were electrospun on the rotating target wheel, removed, and glued to the cover slip at the ends of the fiber bundle with PLGA (85:15). (E, F) Cover slips were painted with a stripe of PLGA down the center and taped to the wheel, and PLLA fibers were electrospun while the PLGA was still wet. Additionally, a “moat” was created in this preparation by painting a wall of PLGA on the perimeter of the cover slip to hold the cell suspension at the time of plating. SEM images reveal that fiber alignment is equal among each substrate design. Cover slips are 22 × 22 mm2, scale bar = 20 μm.