In-House Manufacturing of 3D Culture Chips via Vacuum Thermoforming for Enhanced Imaging Applications

Engineered 3D in vitro cancer models, particularly those that facilitate image-based readouts capable of distinguishing the behavior of different cell populations, have become crucial tools in the discovery process. One such model, 96-GLAnCE (Gels for Live Analysis of Compartmentalized Environments), allows for longitudinal imaging of tumor cell dynamics and therapy response. However, the widespread adoption of 96-GLAnCE has been limited by the need for expensive, specialized fabrication equipment. To overcome this challenge, we have optimized a desktop vacuum thermoforming technique for the in-house production of 96- GLAnCE bottom chips using thin polystyrene films. This optimization has led to the reliable and consistent fabrication of devices. Notably, using thin polystyrene films reduces the overall thickness of the chips, enabling high-magnification imaging for studying primary tumor cell phenotypes in 3D with single-cell and subcellular resolution. Our thermoformed devices offer a flexible, cost-effective solution for addressing a wide range of biological questions across various time scales.