How to orient cells in micro-cavities for high resolution imaging of cytokinesis and lumen formation

TL;DR

This paper introduces a 3D microfabrication technique that precisely orients cells near the microscope coverglass, enhancing high-resolution imaging of cellular processes like cytokinesis and lumen formation in 3D cultures.

Contribution

A novel microfabrication method for positioning and orienting cells close to the coverglass to improve imaging resolution in 3D cellular studies.

Findings

Improved spatial resolution in microscopy, including super-resolution STED.

Effective orientation of cells for studying cytokinesis and lumen formation.

Potential to advance understanding of cell self-organization.

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.