Computational protocol: Angiotensin II Induced Cardiac Dysfunction on a Chip

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Protocol publication

[…] Muscular thin films were placed in warm Tyrode’s solution and the temperature was allowed to briefly fall below 32°C to facilitate the phase transition of the sacrificial poly(NIPAAm) (Polysciences, Inc., Warrington, PA) layer to an aqueous phase allowing for easy release of the PDMS film from the glass base. Once the poly(NIPAAm) reached aqueous phase, films were gently peeled from the glass and tissues were paced at 2 Hz using a Myopacer field stimulator (IonOptix Corp., Milton, MA). The voltage was adjusted to 20% above the minimum voltage required to capture 90% of the total MTFs. Videos were recorded using a Basler camera at 100 frames per second for 3 seconds using Lab View (National Instruments, Woburn, MA) and imaged using a Zeiss Discovery V8 Stereo microscope (Oberkochen, Germany). All experiments were performed at 36 ± 1°C. Custom ImageJ and Matlab codes were created to calculate peak systolic, diastolic, and twitch stress generation using the film radius of curvature, measured PDMS film thickness, and cell layer thickness as inputs into a modified Stoney’s equation []. [...] A student’s t-test was performed with Microsoft Excel® (Microsoft, Redmond, WA) for cell shape, sarcomere organization, tissue thickness, calcium, and muscular thin film studies. P values of less than 0.05 or 0.01 were considered statistical significant and are noted appropriately. Two-way ANOVA analysis was performed with SigmaPlot (Systat Software, Inc., San Jose, CA) to determine statistical significance relative to control tissues for gene expression studies. Genes with p values less than or equal to 0.01 or less than 0.05 were considered significant and are also indicated appropriately. […]

Pipeline specifications

Software tools ImageJ, SigmaPlot
Applications Miscellaneous, Microscopic phenotype analysis
Diseases Heart Diseases
Chemicals Angiotensin II, Calcium