Abstract Body: The study of dynamic Ca2+changes in in vitro models of cardiomyocytes has become a cornerstone to understand the role of calcium signaling in healthy and diseased hearts1. Atomic force microscopy (AFM) is a highly-sensitive and versatile method that can be integrated with optical microscopy and calcium imaging2. We have implemented a combined set up to measure contractilityand calcium waves in human cardiac models. We successfully report the first simultaneous recording of cardiac contractility, though AFM mechanocardiograms (MCG)3 and local calcium waves probed on embryoid bodies. Ensemble empirical mode decomposition (EEMD) filtering was proved the best in terms of signal-to-noise ratio and signal distortions. Caffeine stimulation confirmed the detection capabilities of the used algorithms, measuring the expected physiological response (e.g. calcium duration increased 5.5% and contraction increased 14.5%). The combination of AFM and calcium imaging allows accurate analysis of complex excitation contraction coupling during physiological, disease and drug-induced situations.Figure. A) AFM contraction profiles B) Relative Calcium transients. t0=signal onset; t100=signal offset; tmax=amplitude; t50I=time to rise 50%; t50r=time to fall50%. References:1. Herron TJ, Lee P, Jalife JJ. Circ Res 110, 2012. 2. Pesl M, Pribyl J et al., J Mol Recognit e2602, 2016. 3. Pesl M, Pribyl J et al., Biosens Bioelectron 85, 2016.