Innovative nano-electrode chip for electroporation in yeast

Electroporation is a widely used technique for gene transfection, protein expression, and cellular labeling, but conventional electroporation approaches typically require high-voltage pulses that induce Joule heating, reduce cell viability, and yield heterogeneous efficiency at the single-cell level. These challenges are exacerbated in fungi such as Saccharomyces cerevisiae , whose rigid yeast cell wall acts as an additional barrier to molecular delivery. In this study, we developed a low-voltage yeast transformation strategy using a three-dimensional titanium nitride nano-electrode array fabricated via customized CMOS processes. The nano-scale architecture enhances local electric field strength, while the array design expands the effective manipulation area. Integrated with a data acquisition system, the platform provides programmable control of voltage, frequency, and waveform, enabling semi-automated operation. By combining dielectrophoresis for yeast positioning with electroporation, yeasts were first captured at the electrode sites and subsequently permeabilized for intracellular delivery of propidium iodide. This chip-based system not only concentrates yeasts within the effective field region but also achieves efficient electroporation under reduced voltage conditions, thereby minimizing cellular damage and improving biocompatibility. Using this approach, a transformation efficiency of 52.54 ± 23.25 % with a yeast viability of 58.9 ± 21.5 % was achieved with significantly lower dye concentrations compared to conventional methods. These results demonstrate the feasibility of CMOS-based 3D nano-electrode chips as a resource-efficient, scalable, and biocompatible platform for yeast transformation, highlighting their potential as an enabling technology for future biotechnological and synthetic biology applications.