eLabNotebook > Bioassays > Cytokine ELISA 1-3 Plates Biomek® 2000

Automation of 1-3 Plates of a Beckman Coulter-Immunotech Cytokine ELISA for IL-8 on the BiomekŪ 2000 Workstation

V. Rocchi, Ph. Denizot  and J. Fieschi
Immunoanalysis Department, Research & Development
Immunotech, a Beckman Coulter Company

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Problem Possible Cause Solution
The bottom of the wells of the assay plate is deeply scraped.
  • The height of the needle during washing steps is too low.
  • Check the Z-axis alignment or increase the height value of the wash steps (not above 5%).
Frequent crashes happen at Tip Racks due to the used tips that do not go back properly into the rack.
  • The material of the tips is not compatible with the ELISA kit reagents.
  • Prefer Beckman-Coulter Tips (P/N 372654) that were used during the validation process.
B2K crashes when attempting to pipette samples.
  • Samples are not in the same labware as specified in the method.
  • Go to edit, open labware and check the "BCIOT sample" format. Overwrite the entry with parameters from the actual source plate.
The Wash 8 Tool purge reservoir in the tool rack overflows during a purge.
  • The vacuum pump is not turned on. 
  • Turn on the vacuum pump. 
  • Wash 8 Tool is not aspirating.
  • Possible leak in the aspirate plumbing or vacuum pinch valve of wash unit is not operating properly.
The OD background is over 0.5 Units at 450nm and/or the CVs of duplicates are abnormally high.
  • Wash 8 Tool problem.
  • Check the wash tool needles for salt depots. Check the gasket and change it when needed.
  • Improper set up of 6 port valve.
  • Check the 6 port valve set up in the method and connect the port tubing accordingly.
  • Improper aspiration of reagents.
  • Check that the vacuum pressure is about 13"Hg.
  • Improper distribution of reagents.
  • Check the liquid level in the tips visually. If a difference is visible: Check the reagent for clots or large crystals.
  • Check the correct fixing of the tips on the pipette tool.
  • Check the Worksurface and Side extensions alignment.

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