In the case of SH4-HASPB-GFP, ∼97% of the negative controls did not return scores above a threshold of 2.5. In the case of SH4-YES1-Cherry, 98% of the negative controls were below a threshold of 3.5. Based on these data, scores of 2.5 for SH4-HASPB-GFP and 3.5 for YES1-mCherry were chosen as suitable thresholds for initial identification of potential phenotypes.