Objective

Drug discovery research increasingly utilizes 3D organoid and spheroid cell culture due to their increased reliability as estimators for in vivo efficacy and toxicity [1]. High cellular density and large size relative to 2D cell culture, however, make automated image analysis accompanying microscopy-based screening more difficult. By applying robust algorithms to combined brightfield and multicolor fluorescence imaging, we have created software to morphometrically characterize each spheroid in multiple channels. Our approach accesses information about multiple cell types within each culture system, identifying spatial distributions of individual, categorized cells. We utilize widefield microscopy to maintain the fast acquisition and high throughput capacity required of large drug screening trials. Our analyses provides multi-parametric growth curves, which identify differences in growth patterns to describe perturbed physiology in addition to cytotoxicity in multiplexed assays.

[1] Edmondson, R, Broglie, JJ, Adcock, AF, Yang, L. “Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors” Assay Drug Dev Technol. (2014), 12, 207-218, DOI: 10.1089/adt.2014.573.