A fluorescence-based opto-mechatronic screening module (OMSM) for automated 3D cell culture workflows

Abstract

Cost-effective automated solutions for hydrogel-based 3D cell culture workflows are required to increase throughput, reproducibility and user-independent results. 3D bioprinters can handle viscous biomaterials, however, throughput is low, they are limited to specific biomaterials, and lack post-processing and analysis methods. Conversely, liquid handling robots have higher throughput but miss viscous materials handling capabilities. Furthermore, commercial systems are not only expensive but mostly not ‘open-source’ and hence are not adaptable to specific experimental requirements.The aim of this study was the implementation of an opto-mechatronic screening module (OMSM) for automated 3D cell culture workflows from production to analysis into a previously developed biomanufacturing workstation. An analysis module with a fluorescence wide-field microscope and a motorised XYZ stage was engineered to transport tissue-culture plates from the storage rack of the workstation to the custom-made microscope and to assess fluorescence-based cell parameters. The microscope has two fluorescence channels and a resolution greater than 228 lp/mm. The XYZ-stage achieved an accuracy of 0.8 μm in X-direction (repeatability 2.4 μm) and 1.4 μm in Z-direction (repeatability <0.1 μm). Viability assays and drug-response studies were conducted on 2D and 3D prostate cancer gelatin-methacryloyl hydrogel models demonstrating its capabilities comparable to a conventional fluorescence microscope. Altogether, the integration of an opto-mechatronic screening module into the biomanufacturing workstation enables automated, reproducible, and user-independent production, imaging and analysis of hydrogel-based 3D cell cultures.