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Related Experiment Videos

Location dependent biases in automatic 96-well microplate readers.

R O Harrison1, B D Hammock

  • 1University of California, Department of Entomology, Davis 95616.

Journal - Association of Official Analytical Chemists
|September 1, 1988
PubMed
Summary

Automatic readers used in 96-well microplates can exhibit well location dependent bias, impacting accuracy. A reverse plate wet test effectively identifies these biases, crucial for instrument quality control.

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Area of Science:

  • Biomedical instrumentation
  • Analytical chemistry
  • Laboratory automation

Background:

  • Automated readers in 96-well microplates are essential for quantitative assays.
  • Instrument specifications typically focus on precision and accuracy, often overlooking well location-dependent biases.
  • Such biases can significantly affect experimental results and data reliability.

Purpose of the Study:

  • To investigate well location-dependent biases in automatic microplate readers.
  • To evaluate the effectiveness of a reverse plate wet test for detecting these biases.
  • To assess the performance of 16 different automatic readers against established specifications.

Main Methods:

  • A reverse plate wet test was employed, comparing readings of dye-loaded plates in normal and reversed positions.
  • This method isolates well location bias by analyzing differences in repeated readings, independent of pipetting errors.
  • Sixteen automatic microplate readers from various designs were tested.

Main Results:

  • Significant well location-dependent biases were observed across most of the 16 instruments tested.
  • Only 4 instruments met manufacturer specifications for precision, and only 1 met accuracy specifications.
  • The reverse plate wet test identified severe biases even in instruments with excellent repeatability and passing diagnostic tests.
  • Routine service did not demonstrably improve a reader with significant bias.

Conclusions:

  • Well location-dependent bias is a prevalent issue in automatic microplate readers, affecting accuracy.
  • The reverse plate wet test is a valuable diagnostic tool for identifying and quantifying these biases.
  • Implementing this test during instrument manufacture and routine use is crucial for ensuring reliable quantitative results.

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