Cardio analysis software

Throughout the study, contraction analysis was performed using a high-speed video microscopy followed by motion vector analysis to investigate the contractile characteristics of iPSC-CM monolayers. Depending on the design of the experiment, iPSC-CMs were seeded at 2×10⁵ cells/cm² into 384-well or 96-well plates. Contractility was measured using the Sony SI8000 cell motion imaging system. Video imaging of beating iPSC-CMs was recorded for 10 sec at a frame rate of 75 fps, a resolution of 1024 × 24 pixels, and a depth of 8 bits using a 10× objective on a fully automated Nikon microscope (Eclipse Ti, Nikon). Motion detection and analysis were performed using the Sony Cardio-analysis software (based on a block matching algorithm).^{13 (link)} For stress experiments, iPSC-CMs were treated with the following options: 30 nM triiodothyronine (T3), 1 µM dexamethasone (dex), 20 ng/µl insulin-like growth factor 1 (IGF1) for 72 hr (all Sigma Aldrich); 25 nM digoxin (Sigma Aldrich) for 3 days,1 nM digoxin for 7 days; 10 µM phenylephrine (PE), 25 pg/ml endothelin-1 (ET1), and 100 nM isoproterenol (Iso) for 7 days (all from Sigma Aldrich).