Fig. 2

Comparison of automatically and manually measured cell boundaries for three conditions. a Raw image of merged red/blue channels with superimposed auto-generated cell boundaries (Condition: Chol), arrow identifies sample cell with automated area and centroid coordinates. b Shows manually measured output for one cell, traced three times, showing a range on area estimates, and little change in centroid location, the average area is treated as the manual area estimate for each cell. c Cropped portion of raw image with superimposed automatically (yellow) and manually (red) measured centroid estimates. d Sample of manually traced cell boundaries. (Graph) Corresponding manual area (a^) and automated (a*) are distributed around a 1:1 line of correspondence (161 data points). Absolute percent difference between (a^) and (a*) is on average 33%. Out of 161 manually measured cell boundaries, 95 (59%) area estimates measured below their automated counterpart and 66 (41%) over-estimated the area