Fast quantitative time lapse displacement imaging of endothelial cell invasion

Autoři: Christian Steuwe aff001;  Marie-Mo Vaeyens aff002;  Alvaro Jorge-Peñas aff002;  Célie Cokelaere aff001;  Johan Hofkens aff003;  Maarten B. J. Roeffaers aff001;  Hans Van Oosterwyck aff002
Působiště autorů: Centre for Membrane Separations, Adsorption, Catalysis and Spectroscopy for Sustainable Solutions (cMACS), Department of Microbial and Molecular Systems (MS), KU Leuven, Leuven, Belgium aff001;  Biomechanics Section (BMe), Department of Mechanical Engineering, KU Leuven, Leuven, Belgium aff002;  Molecular Imaging and Photonics, Department of Chemistry, KU Leuven, Leuven, Belgium aff003;  Prometheus, Division of Skeletal Tissue Engineering, KU Leuven, Leuven, Belgium aff004
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article


In order to unravel rapid mechano-chemical feedback mechanisms in sprouting angiogenesis, we combine selective plane illumination microscopy (SPIM) and tailored image registration algorithms — further referred to as SPIM-based displacement microscopy — with an in vitro model of angiogenesis. SPIM successfully tackles the problem of imaging large volumes while upholding the spatial resolution required for the analysis of matrix displacements at a subcellular level. Applied to in vitro angiogenic sprouts, this unique methodological combination relates subcellular activity — minute to second time scale growing and retracting of protrusions — of a multicellular systems to the surrounding matrix deformations with an exceptional temporal resolution of 1 minute for a stack with multiple sprouts simultaneously or every 4 seconds for a single sprout, which is 20 times faster than with a conventional confocal setup. Our study reveals collective but non-synchronised, non-continuous activity of adjacent sprouting cells along with correlations between matrix deformations and protrusion dynamics.

Klíčová slova:

Angiogenesis – Collagens – Deformation – Fluorescence imaging – Fluorescence microscopy – Gels – Light – Optical lenses


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