Figure 6.

Pluri-IQ Performance Evaluation in Different mESC Culture Conditions

(A1) The training set was built by selecting 15 pluripotent, 15 mixed, and 20 differentiated colonies. Manual validation showed that accuracy of the classifier is ∼ 97%. Classifier was updated after manual validation and automatic score was run in five different large images. Manual validation performed afterward showed accuracy values all above 90%. Scale bar, 500 μm (in raw images).

(A2) After selection of a reduced number of colonies to train the classifier, the manual accuracy decreased to ∼59%. However, the update of the classifier after the manual training leads to an increase of accuracy classification.

(B1) Excel sheet output derived from the automatic data comparison.

(B2) Percentage of pluripotent, mixed, and differentiated colonies (left) and area (right) of each condition. A minimum of 100 colonies was analyzed per condition. Color code: green, pluripotent colonies; blue, mixed colonies; red, differentiated colonies.

(C) Normalized mRNA expression fold change (determined by RT-qPCR) of mESC cultured for 4 days in the presence of 2i medium, LIF, 50 nM AA or in the absence of LIF. All samples were analyzed in two technical replicates. All data are presented as means ± SEM. ^∗p < 0.01. AA, Antimycin A; LIF, leukemia inhibitory factor; 2i, serum-free medium with CHIR99021, PD0325901, and LIF.