Fig. 3.

Conceptual figure for the workflow of the quantitative metrics. The metrics based on Fourier transformation were evaluated on the whole image, whereas for the other metrics the automatic Otsu threshold was applied. For histogram, edge and intensity based metrics, the internal circle option was also enabled to assess the information content at the center of the objects only. The results for each metric were visualized as a plot describing sharpness across the spheroid, where the coordinate in the x-axis represents the number of the image within the stack, and the y-axis is the score obtained by the metric. For histogram, edge and intensity based metrics, only the results for the internal circle are represented. Dark blue curves represent the uncleared spheroid, and gray curves represent the cleared spheroid. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)