Image processing procedure to determine the cell count and dispersion values of a well.

<p>(A) Colony nuclei image of each well was obtained by stitching four adjacent, non-overlapping fields together. The example here shows a primary cell colony surrounded by several cell outliers. (B) Nuclei segmentation, which consists of a wavelet transform and watershed algorithm steps, was applied to identify all nuclei in the well. (C) The nuclei segmentation mask was then dilated to generate merging region areas where distinct cell clusters could be isolated. In general, the largest region (yellow), representing the cell colony of interest, and other smaller regions (other colors), representing outlier cell clusters, were identified. (D) Nuclei within the colony of interest were kept for measurement. Cell count was determined by the total nuclei count within the colony. Cell dispersion was determined by applying the spreading coefficient formula. The blue arrow represents a vector centered on the colony center with distance equal to the spreading coefficient.