Additional file 9: Figure S6. of Long-term microfluidic tracking of coccoid cyanobacterial cells reveals robust control of division timing

Growth behavior is similar across all chambers in light-dark cycle experiment. (a) Total growth in different chambers under light-dark cycles. Substantial growth is observed during illuminated periods across all microfluidic chambers. In the dark, minimal growth is detected. (b) Residual errors (gray, with mean shown in black) of exponential fits to lineage growth curves during the illumination periods L1 and L2 of Fig. 3a. (PDF 366 kb

Scalable Device for Automated Microbial Electroporation in a Digital Microfluidic Platform

Electrowetting-on-dielectric (EWD) digital microfluidic laboratory-on-a-chip platforms demonstrate excellent performance in automating labor-intensive protocols. When coupled with an on-chip electroporation capability, these systems hold promise for streamlining cumbersome processes such as multiplex automated genome engineering (MAGE). We integrated a single Ti:Au electroporation electrode into an otherwise standard parallel-plate EWD geometry to enable high-efficiency transformation of Escherichia coli with reporter plasmid DNA in a 200 nL droplet. Test devices exhibited robust operation with more than 10 transformation experiments performed per device without cross-contamination or failure. Despite intrinsic electric-field nonuniformity present in the EP/EWD device, the peak on-chip transformation efficiency was measured to be 8.6 ± 1.0 × 10⁸ cfu·μg^–1 for an average applied electric field strength of 2.25 ± 0.50 kV·mm^–1. Cell survival and transformation fractions at this electroporation pulse strength were found to be 1.5 ± 0.3 and 2.3 ± 0.1%, respectively. Our work expands the EWD toolkit to include on-chip microbial electroporation and opens the possibility of scaling advanced genome engineering methods, like MAGE, into the submicroliter regime