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Updated: Jan 15, 2026
Three-Dimensional 3D Tumor Spheroid Invasion Assay
Published on: May 1, 2015
Protocol for AI-assisted quantitative analysis and setup of tumor spheroid invasion into tissue
Celestine Z Ho1, Daniella B Lawther2, Christina Bao-Xian Huang2
1Mechanobiology Institute, National University of Singapore, Singapore, Singapore; Department of Biomedical Engineering, National University of Singapore, Singapore, Singapore.
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Recent work has revealed an active mechanical role for the tissue barrier in collective invasion, offering a new perspective on tissue invasion and morphogenesis. Here, we present a protocol for AI-assisted quantitative analysis and setup of tumor spheroid invasion into mesothelial tissue. We describe steps for co-culture setup, time-lapse imaging, AI training, and segmentation. We then detail procedures for cell tracking and quantitative analysis to study the fracturing process of the mesothelium. For complete details on the use and execution of this protocol, please refer to Wu et al.1.
