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Minimizing Edge Effect in 96-Well Plates Using CytoOne®

Cells grown in 96-well plates often overlap towards the outer edges of the wells. This tendency, commonly called "edge effect" can be minimized using CytoOne. CytoOne® cell culture plates have a complete 360° Protection Perimeter™ around each well. Filling these inter-well spaces with DMEM media during incubation helps produce a more uniform cell layer.


Common 96-Well Plate Edge Effect
Reduced edge effect in CytoOne

Reduced Edge Effect in CytoOne®
Reduced edge effect in CytoOne


Both plates were seeded and incubated under the same conditions*. The plate on the right shows the growth after filling the inter-well spaces. The edge effect becomes visible after staining.


To decrease the edge effect, fill the inter-well spaces of a CytoOne® 96-well plate with 6 ml of sterile serum-free media (DMEM or similar liquid). Use a 1 ml pipet tip to slowly dispense media around the wells. A slight agitation of the plate may be required to facilitate the spreading of media around the wells. DMEM is recommended instead of H2O as it has a lower surface tension and flows more easily around each well. Do not use surfactants or detergents to lower the surface tension of the liquid, as they could damage the cells if accidentally splashed into the growth wells.

*NIH3T3 cells stained with Crystal-Violet after 24 hours at 37°C, 5% CO2. Initial seeding of 0.01 x 106 cells per well.

See more about CytoOne TC plates.