Journal: Journal of Molecular and Cellular Cardiology Plus
Article Title: Pericellular oxygen dynamics in human cardiac fibroblasts and iPSC-cardiomyocytes in high-throughput plates: insights from experiments and modeling ☆
doi: 10.1016/j.jmccpl.2025.100464
Figure Lengend Snippet: Overview of experiments and modeling of pericellular oxygen in HT 96-well plates. A. The experimental system and the experimental conditions are shown. In-incubator-operated optical imaging system included a camera, a lens, LED ring, and bellows extension to accommodate a glass-bottom 96-well plate. The ratiometric optical readout was calibrated in values of pericellular oxygen concentration over time. The experimental conditions included two cell types - adult cardiac fibroblasts, cFBs, and human iPSC-CMs, three cell densities, and four different culture medium volumes. B. Confirmation of cell growth on the optical oxygen sensors was done using a two-photon upright microscope with a water-immersion lens. Images of hiPSC-CMs on top of the sensor are shown at two different magnifications (25× and 40×). Color encodes depth in the lower magnification image, in which cells are labeled with Hoechst for nuclei and with ChR2-eYFP to highlight the membrane. The higher magnification image shows cytoskeleton (α-actinin) and nuclei (Hoechst); scale bar for the zoomed in version is 50 μm. C. The computational modeling framework is a combination of 3D physics-based simulations of reaction-diffusion in the wells of a HT plate and optimization algorithms to derive empirically-based models of pericellular dynamics in human cFBs and iPSC-CMs, grown in glass-bottom 96-well plates, for a variety of cases. Simulations and data processing were done in Matlab, LiveLink and Comsol.
Article Snippet: The use of LiveLink allows for the workflow shown in C. This feature allows for the scripting of COMSOL with MATLAB and automates the preprocessing and postprocessing steps required for parameter estimation and curve fitting.
Techniques: Optical Imaging, Concentration Assay, Microscopy, Labeling, Membrane, Diffusion-based Assay
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