DIB‐BOT: An Open‐Source Hardware Approach for Plate‐Integrated Droplet Interface Bilayer Deposition

Abstract Droplet interface bilayers (DIBs) offer a controlled lipid environment for studying membrane‐bound processes, with applications in artificial cells, biosensing, and biophysics. Current DIB fabrication faces challenges due to time‐consuming processes and specialized equipment, limiting scale‐up and hindering statistical significance in single‐molecule assays. This research introduces “DIB‐BOT,” an open‐source solution combining a nanoinjector and a 3D printer. DIB‐BOT enables rapid, reproducible DIB fabrication, overcoming manual limitations. Using off‐the‐shelf components, DIB‐BOT ensures high spatial reproducibility, minimal user input, and scalable experiments. The system's utility is demonstrated through pairwise droplet assembly and a fluorescence plate‐reader assay. Compared to manual fabrication, DIB‐BOT shows a 10‐fold reduction in droplet volume error, a threefold reduction in positional error, and 100% droplet yield. This method lowers entry barriers to DIB research, expanding its applications and enhancing data quality.