Analysis of cellular composition of co-culture spheroids

Staining efficiencies were higher than 95% as quantified in preliminary experiments with 2D cultures. Depending on the staining procedure, variations from uniform to punctate signals were detected. The size of all co-culture spheroids decreased over time and snap-freezing did not lead to shrinkage of the spheroids. We were able to detect organizational patterns of different cell types within the spheroids. It was possible to determine the cellular composition by quantitative microscopic analyses of cryo-sections as it could be confirmed by flow cytometric analyses. Depending on the experimental requirements, a combination of both methods might lead to valuable synergy.