Liquid Handling · Automated pipetting · Imaging integration · Microscopy integration · Workflow automation · 2025 · bioRxiv
Open-source cell culture automation system with integrated cell counting for passaging microplate cultures
Tissue culture in 96-well microplates is conventionally a tedious, highly manual process sensitive to individual technique and experimenter error. Here, we describe the Automated Cell Culture Splitter (ACCS), a system for passaging plates of adherent or suspension cells, for routine culture maintenance or specialized applications such as seeding plates for microscopy.
Performance the paper reports
2 of 5- Motion—Motion accuracy: not reported
- Volume10%Volumetric accuracy: reported
- Viability—Cell viability: not reported
- Unattendedwalk-awayUnattended operation: reported
- Standard—Standard compliance: not reported
- Volumetric accuracy10%
The useful lower bound is dictated by the acceptable error for the application; for example, the best-case statistically achiev- able CV is ∼10% at a sample concentration around 100 k cells/mL and gets worse with decreasing concentration.
- Unattended operationwalk-away
While the end-to-end time for a run, including the setup, teardown, and robot operation, is significantly longer than for the manual pro- cess, there is a substantial period of “walk-away” time during which the operator is able to attend to
How hard this is to adopt
3.4 of 5, higher is easierEach score is shown against the range the other 59 builds actually occupy, because no criterion uses the full 1–5 scale. These describe how reproducible the build is, not how good the science is.
Resolution
2/1–5P300 pipettes handling 50 uL trypsin, 100 uL DPBS and 100-200 uL seeding; aspiration above 180 uL causes bubbles; seeding CV under 11%.
Throughput
4/2–5P300 8-channel head plus an independent single-channel pipette on separate mounts and an 8-chamber counting flow cell, 2-3 plates/day; per-well seeding is still serial.
Parts sourcing
3/2–4Mix of common parts and custom/printed components.
Skill needed
4/2–4Build/operation described as low-skill or basic assembly.
Facilities needed
4/2–4Uses common benchtop/desktop equipment or generic consumables.
Documentation
4/2–5Open resources include several build or operation artifacts.
Validation effort
4/2–4Cell counter benchmarked against a flow cytometer over 100-400k cells/mL with 32 reads/sample; ATP assay versus manual seeding gave 54% lower CV and 92% vs 52% imaging keeper rate.
Cycle time
2/1–4Hands-on time cut 61% (under 30 min vs an hour or more), but the end-to-end run of under 3 h is roughly twice the manual duration.
2 further criteria are recorded but not shown
- Application Level — scored 4 (75% of records share one value), so it cannot separate one build from another.
- Build Time (scored for only 6 of 60 records), so it cannot separate one build from another.
Both remain in the published dataset.
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Matched by doi · retrieved 2026-07-21
Abstract
Tissue culture in 96-well microplates is conventionally a tedious, highly manual process sensitive to individual technique and experimenter error. Here, we describe the Automated Cell Culture Splitter (ACCS), a system for passaging plates of adherent or suspension cells, for routine culture maintenance or specialized applications such as seeding plates for microscopy. The system is built around the Opentrons OT-2 liquid handling robot and incorporates a novel on-deck imaging-based cell counter which allows it to compensate for density disparities across a source plate and control the number of cells seeded on a per-well basis. We find this solution can cut hands-on time by 61% and the results compare favorably to our existing manual cell culture processes in terms of both seeding density precision and bio-logical outcomes, achieving a control of seeding density with a well-to-well coefficient of variation (CV) under 11%. The system is designed to be adaptable and an accessible entry point into automation for high-throughput cell culture; to that end, all of the source code and hardware designs are released under open source licenses.
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