Polydimethylsiloxane (PDMS)-based neuron microengineering provides new opportunities for spatiotemporal control of neuronal activity and stimuli. The demand for long-lasting adhesive PDMS surfaces has steered the development of straightforward, feasible, and accessible interface modifications. Here, we describe an innovative approach for promoting and engineering neuron adhesion on a PDMS substrate based on a very simple modification using poly-d-lysine-conjugated Pluronic F127, a functionalized triblock polymer. The modification procedure only involves single-step pipetting or microfluidic-guided introduction for the reinforcement of cell adhesion in quantity, extensibility, and stability. Micropatterning at a single-cell resolution, microfluidic long-term culture, and neuron network formation were achieved. The present approach provides a previously unprecedented simple and effective technique for neuron adhesion on PDMS and may be useful for applications in neurobiology, tissue engineering, and neuronal microsystems.