Common issues & fixes
| Symptom | Likely cause | Fix |
|---|---|---|
| High background across the plate | Insufficient washing or blocking | Increase wash cycles; extend blocking incubation; verify wash-buffer reservoir isn't empty. |
| Weak or flat signal | Reagent expired or under-dispensed | Check reagent volumes and placement; confirm detection antibody concentration. |
| Edge-well variability | Evaporation during incubation | Use a plate lid/seal during long incubations; keep the deck away from airflow. |
| Run paused mid-protocol | Vision flagged a tip/plate exception | Re-seat the flagged item as prompted; the robot resumes from where it stopped. |
When not to automate this
If you run ELISAs only occasionally — a few plates a month — the setup time may outweigh the time saved; manual may still be the right call. And if your protocol changes substantially every run, stabilise it manually first, then automate. Automation amplifies a good process; it won't rescue an unstable one.
Verification note: this protocol is vision-verified on RoR hardware. Still validate against your own positive/negative controls on first run before relying on it for critical samples.