As microfluidics has been applied extensively in many cell and biochemical applications, monitoring the related processes is\nan important requirement. In this work, we design and fabricate a high-throughput microfluidic device which contains 32\nmicrochambers to perform automated parallel microfluidic operations and monitoring on an automated stage of a microscope.\nImages are captured atmultiple spots on the device during the operations formonitoring samples in microchambers in parallel; yet\nthe device positions may vary at different time points throughout operations as the device moves back and forth on a motorized\nmicroscopic stage. Here, we report an image-based positioning strategy to realign the chamber position before every recording\nof microscopic image. We fabricate alignment marks at defined locations next to the chambers in the microfluidic device as\nreference positions. We also develop image processing algorithms to recognize the chamber positions in real-time, followed by\nrealigning the chambers to their preset positions in the captured images.We perform experiments to validate and characterize the\ndevice functionality and the automated realignment operation. Together, this microfluidic realignment strategy can be a platform\ntechnology to achieve precise positioning of multiple chambers for general microfluidic applications requiring long-term parallel\nmonitoring of cell and biochemical activities.
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