Hi,

could you maybe share a picture of your cells?

It is possible, depending on how you add your treatment, that you use too much force when pipetting. so you wash away the cells. Have you considered this?

Best,

Franziska

Hey Prachi,

This sounds like a pretty classic case of edge effect in 96-well plates. A few general things you could try:

Hope this helps!

Best,

Wyatt

Mayo Clinic – Rochester

ive also noticed to prevent edge effect, that if you can let your plate sit (immediately after seeding) inside the BSC for 30 minutes, i.e. to let enough time pass to allow your cells to attach to the bottom of the well, this can drastically promote a more uniform density across wells in my experience.

there is always more cells on the edge than in the center but a more uniforme repartition can be obtained if you seed you cell in a larger volume 200ul To change the medium you can (depending of your experiment) turn the plate over and tap it on layer of absorbent paper (quicker and safer than vacuum...)

Thank you everyone for your insightful answers.

On the day I seed the cells, they appear to be uniformly distributed, and the initial growth also looks even. However, by the next day, when I discard the media—from the edges—I notice that the cells in the center are no longer present.

Nevertheless, I will try to incorporate your suggestions during the next seeding to improve consistency.

Thank you once again for your help.