Figure 4.

Images Showing the Workflow for Manually Quantifying Spheroid Images

(A) Brightfield image of spheroid and emigrated cells.

(B) A maximum intensity projection was created.

(C) The area of an elipse encircling the migration was calculated. The "Find Edges" function was used as needed to improve the signal-to-noise ratio.

(D) Overlap of brightfield and flouresence imaging of nucloefected cells.

(E) Fluorescence imaging of nucleofected cells.

(F) For manual analysis of nucleofected fluorescent cells, the average distance of the eight maximally migrated cells was measured. Scale bar, 100 microns.