Transfection efficiency and viability (relative metabolic activity) in GB 319 cells assessed by (A) flow cytometry and MTS assay and (B) fluorescence microscopy (GFP channel alone is shown). Transfection was carried out with the plasmid released from multilayer films and collected over each time period: 0–1 hr, 1–6 hrs, 6–12 hrs, and 12–18 hrs and combined with polymer 447. Transfection was measured 48 hrs post-transfection. For all, the number in the upper right-hand corner is the final concentration of DNA added to each well in that condition. For all, scale bar: 500 μm.