We use a microfluidic approach to fabricate gelatin fibers with controlled sizes and cross-sections. Uniform gelatin microfibers with various morphologies and cross-sections (round and square) are fabricated by increasing the gelatin concentration of the core solution from 8% to 12%. Moreover, the increase of gelatin concentration greatly improves the mechanical properties of gelatin fibers; the Young's modulus and tensile stress at break of gelatin (12%) fibers are raised about 2.2 and 1.9 times as those of gelatin (8%) fibers. The COMSOL simulations indicate that the sizes and cross-sections of the gelatin fibers can be tuned by using a microfluidic device with four-chevron grooves. The experimental results demonstrate that the decrease of the sheath-to-core flow-rate ratio from 150 : 1 to 30 : 1 can increase the aspect ratio and size of ribbon-shaped fibers from 35 μm × 60 μm to 47 μm × 282 μm, which is consistent with the simulation results. The increased size and shape evolution of the cross-section can not only strengthen the Young's modulus and tensile stress at break, but also significantly enhance the tensile strain at break.