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. 2022 Aug 5;3(3):101602. doi: 10.1016/j.xpro.2022.101602

Figure 1.

Figure 1

High-content screening (HCS) of mitochondrial membrane potential (ΔΨm) in iPSC-derived neural progenitor cells (NPCs)

(A) Schematic of the HCS protocol steps, including the mask established for the detection of ΔΨm in live-cells.

(B) Flow of imaging analysis using CellProfiler software. Scale bar: 100 µm.

(C) Flow of imaging analysis using Columbus software. Scale bar: 100 µm.

(D) ΔΨm intensity evaluated using the CellProfiler image analysis pipeline. Robustness of the HCS protocol was calculated based on Z′ score (for effective high-throughput assays, values should be between 0.5 and 1). Mean ± standard deviation (SD), unpaired Student’s t-test, ∗∗∗∗p<0.0001.

(E) ΔΨm intensity evaluated using the Columbus image analysis pipeline. Mean ± SD, unpaired Student’s t-test, ∗∗∗∗p < 0.0001.

(F) Compound screening performed in NPCs differentiated from human iPSCs based on the HCS protocol described here. Blue dots indicate the positive control samples treated with FCCP and antimycin A (AA). These drugs are known to decrease ΔΨm and their values are considered as 100% inhibition of ΔΨm. Orange dots indicate screened compounds that can lead to 50%–100% inhibition of ΔΨm.