Seeding and initial selection: (A) Open the last image of the time-series to segment and track.

<p>An example cell is selected to build the initial seed (red arrow). (<b>B</b>) A watershed algorithm is applied to the image using a manually set threshold between 0 and 255 (threshold used here = 65). (<b>C</b>) Defects in the watershed are corrected manually. Subsections of the cell may be joined as indicated by the yellow arrow, wheras non-cell material may be removed as indicated by the pink arrow. Compare with the boxed region in (B). (<b>D</b>) The program automatically fills holes and one-pixel ‘cracks’ to complete the seed. (<b>E</b>) Final image for the entire field of view with 16 seeded cells inducated by their color.