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Updated: Feb 16, 2026

In Vitro Assessment of Cardiac Function Using Skinned Cardiomyocytes
Published on: June 22, 2020
MUSCLEMOTION: A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In
Luca Sala1, Berend J van Meer1, Leon G J Tertoolen1
1From the Department of Anatomy and Embryology, Leiden University Medical Center, The Netherlands (L.S., B.J.v.M., L.G.J.T., M.B., R.P.D., M.A.E.D., E.G., C.G., M.R.M.J., M.P.H.M., V.V.O., R.P., C.L.M.); Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary, and Life Science, University of Glasgow, United Kingdom (Q.L., G.L.S., F.L.B.); Hubrecht Institute - Royal Netherlands Academy of Arts and Sciences, Utrecht, The Netherlands (J.B., S.M.K., C.D.K.); Department of Stem Cell Biology, University of Nottingham, University Park, Nottingham, United Kingdom (C.D., D.M.); Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg Eppendorf, Germany (T.E., A.H., I.M., U.S.); DZHK (German Centre for Cardiovascular Research), Partner Site Hamburg/Kiel/Lübeck (T.E., A.H., I.M., U.S.); Department of Experimental Pharmacology and Toxicology, University Medical Center Hamburg Eppendorf, Hamburg, Germany (U.S.); Hart Long Centrum, Leiden University Medical Center, The Netherlands (E.R.H., M.R.M.J.); Department of Applied Stem Cell Technologies, University of Twente, Enschede, The Netherlands (R.P., M.C.R., C.L.M.).; Division of Heart and Lungs, Department of Medical Physiology, University Medical Center Utrecht, The Netherlands (J.B., S.M.K., C.D.K.); and Clyde Biosciences, Ltd, BioCity Scotland, United Kingdom (G.L.S., F.L.B.).
Rationale:
There are several methods to measure cardiomyocyte and muscle contraction, but these require customized hardware, expensive apparatus, and advanced informatics or can only be used in single experimental models. Consequently, data and techniques have been difficult to reproduce across models and laboratories, analysis is time consuming, and only specialist researchers can quantify data.
Objective:
Here, we describe and validate an automated, open-source software tool (MUSCLEMOTION) adaptable for use with standard laboratory and clinical imaging equipment that enables quantitative analysis of normal cardiac contraction, disease phenotypes, and pharmacological responses.
Methods And Results:
MUSCLEMOTION allowed rapid and easy measurement of movement from high-speed movies in (1) 1-dimensional in vitro models, such as isolated adult and human pluripotent stem cell-derived cardiomyocytes; (2) 2-dimensional in vitro models, such as beating cardiomyocyte monolayers or small clusters of human pluripotent stem cell-derived cardiomyocytes; (3) 3-dimensional multicellular in vitro or in vivo contractile tissues, such as cardiac "organoids," engineered heart tissues, and zebrafish and human hearts. MUSCLEMOTION was effective under different recording conditions (bright-field microscopy with simultaneous patch-clamp recording, phase contrast microscopy, and traction force microscopy). Outcomes were virtually identical to the current gold standards for contraction measurement, such as optical flow, post deflection, edge-detection systems, or manual analyses. Finally, we used the algorithm to quantify contraction in in vitro and in vivo arrhythmia models and to measure pharmacological responses.
Conclusions:
Using a single open-source method for processing video recordings, we obtained reliable pharmacological data and measures of cardiac disease phenotype in experimental cell, animal, and human models.
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