Nuclei deformation reveals pressure distributions in 3D cell clusters
Autoři:
Adele Khavari aff001; Allen Joseph Ehrlicher aff002
Působiště autorů:
Applied Chemistry, Chemistry and Chemical Engineering, Chalmers University of Technology, Göteborg, Sweden
aff001; Department of Bioengineering, McGill University, Montreal, Canada
aff002
Vyšlo v časopise:
PLoS ONE 14(9)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0221753
Souhrn
Measuring pressures within complex multi-cellular environments is critical for studying mechanobiology as these forces trigger diverse biological responses, however, these studies are difficult as a deeply embedded yet well-calibrated probe is required. In this manuscript, we use endogenous cell nuclei as pressure sensors by introducing a fluorescent protein localized to the nucleus and confocal microscopy to measure the individual nuclear volumes in 3D multi-cellular aggregates. We calibrate this measurement of nuclear volume to pressure by quantifying the nuclear volume change as a function of osmotic pressure in isolated 2D culture. Using this technique, we find that in multicellular structures, the nuclear compressive mechanical stresses are on the order of MPa, increase with cell number in the cluster, and that the distribution of stresses is homogenous in spherical cell clusters, but highly asymmetric in oblong clusters. This approach may facilitate quantitative mechanical measurements in complex and extended biological structures both in vitro and in vivo.
Klíčová slova:
Physical sciences – Physics – Classical mechanics – Pressure – Osmotic pressure – High pressure – Mechanical stress – Compression – Research and analysis methods – Imaging techniques – Fluorescence imaging – Microscopy – Light microscopy – Confocal microscopy – Chemical characterization – Optical analysis – Refractive index – Biology and life sciences – Cell biology – Cellular structures and organelles
Zdroje
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