image: Cell Counting Imager (CCI), a custom device that measures cell concentrations in 8 samples at once, to calculate the appropriate volumes of cell suspension needed to seed a specified number of cells in each destination well. The imager is an inverted darkfield microscope featuring a custom flow cell with eight chambers and a motorized linear stage that positions the optical system to image each chamber. The flow cell mounts on top of the CCI and has a row of ports that allow the multichannel pipette on the OT-2 robot to inject samples from one column of the source plate at a time.
Credit: Rafael Gomez-Sjoberg
An automated cell culture system reduces hands-on time and improves seeding consistency in 96-well microplates. Manuel Leonetti, Rafael Gómez-Sjöberg, and colleagues developed the Automated Cell Culture Splitter, an open-source robotic system built around the Opentrons OT-2 liquid handling robot equipped with a custom cell counting imaging instrument. The system automates passaging—the transfer of cells from one culture plate to another when they grow too numerous—of either adherent or suspended cells in 96-well plates, a process conventionally requiring tedious manual work. The authors tested the system with a commonly used cell line known as HEK293T. In direct comparisons, plates seeded by the automated system showed higher consistency in cell density than plates seeded manually using visual estimation. The system reduced hands-on time by 61% compared to manual passaging, though total processing time was longer. In sensitive high-throughput imaging applications, plates seeded by the automated system had a 92% rate of usable imaging sites compared to 52% for the best manually seeded plate. According to the authors, the open-source design and low cost of components (~$18,000) makes this automation approach accessible to research laboratories, potentially reducing both labor demands and work-related musculoskeletal disorders associated with intensive manual pipetting, while increasing experimental reproducibility.
Journal
PNAS Nexus
Article Title
Open-source cell culture automation system with integrated cell counting for passaging microplate cultures
Article Publication Date
30-Dec-2025