Fluorescence microscopy of nucleofection optimizations. (A) Microscopy images of the initial nucleofection optimization. Each well was subjected to a particular proprietary electroporation condition, designated by the serial number overlaid on each picture, and preceded by the number 96-(For example: Well B2 corresponds to 96-EH-100). Wells in columns 1–4 represent 32 different electroporation conditions, evaluating cells nucleofected in proprietary reagent SE. Columns 5–8 repeat the same 32 electroporation conditions in proprietary reagent SF, while columns 9–12 evaluate the 32 conditions in reagent SG. Wells H4, H8, and H12 are controls that contained the respective nucleofection reagents, but were not electroporated. (B) Microscopy of the secondary optimization containing SE only. Microscopy is only shown for 1/3 of the plate, representing each unique electroporation condition. Well H4 is the control well which was not electroporated. (C) Well G2 from initial optimization and H2 from SE optimization showing GFP throughout the cells.