Development of Automated Organoid Tracking Platform for High-Throughput Screening

Organoids are three-dimensional tissues structures that form in vitro, mimicking the native microenvironment. Our group previously developed a robotic platform that can automatically deposit complex three-dimensional vascularized organoids of healthy and tumor-derived tissues in 384 well-plate formats. Tissue-embedded microsensors permit a real-time kinetic measurement of oxygen within organoids on the DynamiX™ table-top platform. The DynamiX™ platform allows for 384 experiments to be conducted in parallel, with a resolution of approximately one hour. However, this introduces the need for automation of the platform as all manual processes highly limit throughout and introduce significant biases.

Organoid movement is sporadic, causing 31% of measurements to be lost due to the inability to probe the sensors. Therefore, we developed an automated software using HSV-filtered contour-based vision processing to monitor organoids in real-time, reducing the scanning time by 30-fold, reaching an accuracy of 98.4%. To that end, we implemented a custom-designed stepper-based radial motorized nosepiece with 31.4µm eighth-step precision. Furthermore, analysis of the experimental data was automized to reduce the labor intensity and bias associated with semi-manual procedures. We developed multiple adaptive MATLAB-based platforms capable of creating concise reports from approximately 27,000 raw measurements through the mapping of three-dimensional matrices and custom algorithms, reducing analysis time by 24-fold. This automated approach is an advancement towards an unbiased high-throughput Organ-on-a-Chip technology.