How to orient cells in microcavities for high resolution imaging of cytokinesis and lumen formation.
Fiche publication
Date publication
janvier 2020
Journal
Methods in cell biology
Auteurs
Membres identifiés du Cancéropôle Est :
Pr RIVELINE Daniel
Tous les auteurs :
Bhat A, Lu L, Wang CH, Lo Vecchio S, Maraspini R, Honigmann A, Riveline D
Lien Pubmed
Résumé
Imaging dynamics of cellular morphogenesis with high spatial-temporal resolution in 3D is challenging, due to the low spatial resolution along the optical axis and photo-toxicity. However, some cellular structures are planar and hence 2D imaging should be sufficient, provided that the structure of interest can be oriented with respect to the optical axis of the microscope. Here, we report a 3D microfabrication method which positions and orients cell divisions very close to the microscope coverglass. We use this approach to study cytokinesis in fission yeasts and polarization to lumen formation in mammalian epithelial cells. We show that this method improves spatial resolution on range of common microscopies, including super-resolution STED. Altogether, this method could shed new lights on self-organization phenomena in single cells and 3D cell culture systems.
Mots clés
Biological physics, Cell polarization, Cytokinesis, Microfabrication, Organoids, Super-resolution STED
Référence
Methods Cell Biol.. 2020 ;158:25-41