Fig. 3 | Nature Communications

Fig. 3

From: Simultaneous measurement of excitation-contraction coupling parameters identifies mechanisms underlying contractile responses of hiPSC-derived cardiomyocytes

Fig. 3

Triple Transient Measurement system for determining AP, Ca and Co simultaneously. a Hardware setup: the hiPSC-CM model of interest (2D or 3D) is incubated with the voltage sensitive dye (ANNINE-6plus), calcium sensitive dye (Rhod 3) and cell membrane label (CellMask Deep Red) used for measuring contraction. The hiPSC-CMs are excited using three rapidly alternating LEDs which are synchronized with a high-speed camera. The camera receives the emitted signal from the hiPSC-CMs after photon amplification by the image intensifier. The data are then fed into the data processing pipeline. b The video frames are split into three stacks that contain the information regarding the AP (blue), Ca (yellow) and Co (red). The information of each stack is extracted by either plotting the intensity changes and/or applying MUSCLEMOTION. c The data are then “cleaned” to remove the motion artefacts, invert the signal when necessary, calculate ∆F/F, apply a minimal filter and/or average the events within one recording. d The data are quantified by summarizing the kinetic parameters of interest determined in Table 1. e Finally, the data are plotted and meanwhile fed into the hypothesis algorithm

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