期刊
PLOS ONE
卷 14, 期 9, 页码 -出版社
PUBLIC LIBRARY SCIENCE
DOI: 10.1371/journal.pone.0221753
关键词
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资金
- Chalmers University of Technology, Department of Chemistry and Chemical Engineering
- NSERC (Natural Science and Engineering Research Council of Canada) [RGPIN/05843-2014, EQPEQ/4723392015]
- CIHR (Canadian Institutes of Health Research) [143327]
- Canadian Cancer Society [703930]
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.
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