Peter van Mourik

110 Chapter 4 Table 7. Continued. FIS ASSAY VALIDATION - WORKFLOW & TIMELINES (SUGGESTED) Wk nr Action Extra comments → Follow the organoid disruption & clean up steps. → Seed 64 wells (in a 96 well plate) per reference line and perform FIS assay experiment n1 → Seed 8 wells (in a 24 well plate) per reference line for FIS assay experiment n=2 in wk 6. Wk 6 Take up 8 full wells: → Follow the organoid disruption & clean up steps. → Seed 64 wells (in a 96 well plate) per reference line and perform FIS assay experiment n2 → Seed 8 wells (in a 24 well plate) per reference line for FIS assay experiment n=3 in wk 7. Wk 7 Take up 8 full wells: → Follow the organoid disruption & clean up steps. → Seed 64 wells (in a 96 well plate) per reference line and perform FIS assay experiment n3 → Seed 4 wells (in a 24 well plate) per reference line for back up. Wk 7- 8 Data analysis: ● FIS data should be comparable to Dekkers et al (2016). → Average data of FIS experiments n1, n2 and n3 of all reference lines into n=3 → Compare data to FIS results as published. → If finished, stop organoid cultures.