Super-Resolution-Chip: an in-vitro platform that enables super-resolution microscopy of co-cultures and 3D systems

Area of Science:

• Biotechnology
• Cell Biology
• Microscopy

Background:

• Organs-on-a-chip platforms mimic human physiology for in vitro cell culture.
• Integrating advanced imaging, like super-resolution microscopy (SRM), with these platforms remains challenging.
• Existing SRM-compatible platforms often lack 3D co-culture capabilities or are complex to use.

Purpose of the Study:

• To develop an integrated organs-on-a-chip platform enabling super-resolution microscopy for complex cellular studies.
• To create a user-friendly chip supporting 2D and 3D co-cultures with high-resolution imaging from both sides of a membrane.
• To investigate molecular changes in co-cultured endothelial and epithelial cells using the developed platform.

Main Methods:

• Developed the Super-Res-Chip: a 3D-printed device with a porous membrane for cell co-culture.
• Utilized direct stochastic optical reconstruction microscopy (dSTORM) for molecular imaging.
• Co-cultured endothelial and epithelial cells, including glioblastoma cells, in 2D and 3D configurations.

Main Results:

• The Super-Res-Chip successfully enabled super-resolution microscopy (SRM) on both sides of the membrane.
• High spatial resolution (20-30 nm in xy, 10-20 nm direct) was achieved in both 2D and 3D cultures.
• Analysis of Connexin43 distribution showed differences in cluster density but not in number, size, or radii.

Conclusions:

• The Super-Res-Chip platform facilitates easy characterization of complex cellular processes at the molecular scale.
• This technology enhances the capacity for single-molecule resolution imaging of intricate cellular organizations.
• The platform offers improved capabilities for studying cell-cell interactions and molecular dynamics in physiologically relevant 3D models.