Micro-/nano-scale biological ‘soft’ structures have attracted increasing interest in biomedical research, including the study of cell-material interactions. However, most materials of micro-/nano-fabrication are not suitable for biological applications, as they require extensive post-processing or exhibit high mechanical stiffness. On the other hand, soft materials exhibiting high cytocompatibility require long fabrication times with a decreased spatial resolution of features. Thus, a facile fabrication technique of micro-/nano-scale structures of biological soft materials using a cost-effective and high-throughput method is needed. To achieve this, this study proposed a one-step 3D microfabrication method for biological soft materials in cooperation with a light-induced self-focusing photo-polymerization, a controlled oxygen reaction-diffusion, and digital microprinting. For instance, it was anticipated that this microfabrication technique of soft material provides efficient simple 3D scaffold platform that can address the questions of neural mechanobiology studies on the interaction between biological artificial axons bundle and neurons.