Microscopy-based cellular contractility assay for adult, neonatal, and hiPSC cardiomyocytes

• Published in: STAR protocols 2022-03, Vol.3 (1), p.101144, Article 101144
• Main Authors: Scalzo, Sérgio, Mendonça, Carolina A.T. F., Kushmerick, Christopher, Agero, Ubirajara, Guatimosim, Silvia
• Format: Article
• Language: English
• Subjects: Biophysics; Cell Biology; Cell culture; Cell isolation; Cell-based Assays; Humans; Induced Pluripotent Stem Cells - metabolism; Infant, Newborn; Microscopy; Myocytes, Cardiac - metabolism; Protocol; Reproducibility of Results; Single Cell; Stem Cells
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101144

This protocol provides instructions to acquire high-quality cellular contractility data from adult, neonatal, and human induced pluripotent stem cell-derived cardiomyocytes. Contractility parameters are key to unravel mechanisms underlying cardiac pathologies, yet difficulties in acquiring data can compromise measurement accuracy and reproducibility. We provide optimized steps for microscope and camera setup, as well as cellular selection criteria for different cardiomyocyte cell types, aiming to obtain robust and reliable data. Moreover, we use CONTRACTIONWAVE software to analyze and show the optimized results. For complete details on the use and execution of this profile, please refer to Scalzo et al. (2021). [Display omitted] •High-quality contractility data requires specific settings for different cardiac cells•Distinct cell types require specific selection criteria for proper contractility assay•High-quality recordings require adjustments to focus, diaphragm, and lighting•Non-optimized data acquisition settings can influence cell contractility speed This protocol provides instructions to acquire high-quality cellular contractility data from adult, neonatal, and human induced pluripotent stem cell-derived cardiomyocytes. Contractility parameters are key to unravel mechanisms underlying cardiac pathologies, yet difficulties in acquiring data can compromise measurement accuracy and reproducibility. We provide optimized steps for microscope and camera setup, as well as cellular selection criteria for different cardiomyocyte cell types, aiming to obtain robust and reliable data. Moreover, we use CONTRACTIONWAVE software to analyze and show the optimized results.