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In high-throughput experiments where cells are cultured, treated, stained, and imaged in 384-well microplates, I frequently see significant edge effects. For instance, the following plot shows cell count (as measured by CellProfiler from images) for each well on such a plate:

Example of edge effects

How can I reduce these effects? I can try to correct for them statistically, but it would be so much nicer to reduce them at the wet-work stage.

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1 Answer 1

Lundholt et al. describe a very simple trick: let the plates settle in air at room temperature for a couple of hours before placing it in the incubator. We tried it out, and although the color scales are different in the following plots, it is easy to see that it really does help:

Transferred directly to incubator Left out for 1 h Left out for 2 h

(Thanks to Sigrun Gustafsdottir and Kate Madden for testing this.)

I have seen some people "pad" the plate by using only the 308 interior wells and filling the 76 edge wells with only liquid, but I have no data for how well that works.

Allison Tanner from Corning has collected a compendium of techniques to reduce the occurrence of irregular patterns of cell adhesion or “edge effect” in microplates. Brifly summarized:

  1. Avoid bubble formation in the media when seeding cells.
  2. Use cell harvesting methods that foster adequate mixing of the cells in the media and complete dissociation of the monolayer into single cells.
  3. Seed cells at a density that will support attachment and growth.
  4. Allow cell attachment to occur in media of the proper formulation for the particular cell type.
  5. Dispense cells in an appropriate volume of media.
  6. Follow strict aseptic techniques according to respected microbiological practices.
  7. Cultures should be routinely monitored for mycoplasmal contamination.
  8. Minimize entrance to incubators to help reduce fluctuations in the internal environment.
  9. Allow cells to settle in the microplates while the microplates are on the benchtop.
  10. Culture cells in an environment that reduces the formation of static electrical charges.
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