Book Title Intestinal Differentiated Cells Book Subtitle Methods and Protocols ISBN978-1-0716-3075-4 Chapter book Title Automated quantitative analysis of shape features in human epithelial monolayers and spheroids generated from colorectal cancer cells Description for the image named HCT116_Monolayer: HCT116 colorectal cancer cells have been grown as monolayers, then fixed and stained with Alexa 488-phalloidin to visualize actin filaments. Cells were imaged using a SP8 Leica confocal microscope. The image is the Sum projection of 3 independent planes (0.5 um each), exported to .tiff. The image serves as example for single cell analysis using automated packages such as Cellpose 2.0. Description for the image named HCT116_Spheroid: HCT116 colon cancer cells were seeded on low-attachment plates to generate spheroids. This image was acquired after 16h from seeding cells using a widefield Dmi8 Leica microscope. The image can be used to obtain automated cell contours and cell shape features from individual cells forming the spheroid using Cellpose 2.0 package and ImageJ/Fiji. A description of the steps to follow could be found in Brown and Juanes, 2023, ' Automated quantitative analysis of shape features in human epithelial monolayers and spheroids generated from colorectal cancer cells', a chapter in the "Intestinal Differentiated Cells" Book from the Methods in Molecular Biology - Springer Protocols from Nature Humana Press editorial.

Tiff images of HCT116 cell monolayers and 3-Dimensional (3D) spheroids were kindly provided by Lautaro Baro and Asifa Islam, respectively, both members of our lab. Hannah M. Brown and M. Angeles Juanes are the authors of this chapter book. This work was supported by grants from the Academy of Medical Science -the Wellcome Trust/the Government Department of Business, Energy and Industrial Strategy/British Heart Foundation/Diabetes - AMS Springboard Award UK [SBF006\1070], and the CIDEGENT Excellent Research Program from the Valencian regional government CIDEGENT/2021/026 to M.A.J.