How to orient cells in microcavities for high resolution imaging of cytokinesis and lumen formation

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Alka Bhat - , University of Strasbourg (Author)
  • Linjie Lu - , University of Strasbourg (Author)
  • Chen-Ho Wang - , Chair of BioNano-Tools, Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Simon Lo Vecchio - , University of Strasbourg (Author)
  • Riccardo Maraspini - , Chair of Biophysics, Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Alf Honigmann - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Daniel Riveline - , University of Strasbourg (Author)
  • French National Centre for Scientific Research (CNRS)

Abstract

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.

Details

Original languageEnglish
Pages (from-to)25-41
Number of pages17
JournalMethods in cell biology
Volume158
Publication statusPublished - 2020
Peer-reviewedYes

External IDs

Scopus 85081283432
ORCID /0000-0003-0475-3790/work/155291299

Keywords

Keywords

  • Animals, Cytokinesis, Dogs, HeLa Cells, Humans, Imaging, Three-Dimensional/methods, Madin Darby Canine Kidney Cells, Microscopy, Fluorescence, Microtechnology/methods, Polymers/chemistry, Time Factors