Fig. 3
Optimization of the MeioSeed thresholding settings to acquire a fit with the model for crossover frequency described by Melamed-Bessudo et al. [16]. A set of 100 brightfield, RFP and GFP fluorescence stereomicroscopy images collected from ten independent Col3-4/20 (♂) × Ler-0 (♀) F2 populations were run in MeioSeed with different combinations of ‘FilterObjects’ thresholding settings for RFP and GFP fluorescence. Presented are the counted fractions of which are green, red, green and red or non-fluorescent seeds among the total number of seeds, along with the calculated crossover frequencies. The theoretical fractions according to the model are also provided. The best fit with the model was achieved with the ‘FilterObjects’ settings of 0.12 for red fluorescence, 0.077 for green fluorescence and subsequent removal of photos with a ≥ fourfold scewed ratio of red:green fluorescent seeds (or vice versa)