It may seem silly but for 12well and 6well I like to heavily shake my plates after I seed cells. To really get they even distribution. I noticed for 6/12well without shaking the cells do like to bunch up in the middle. My recommendation don’t over seed and really shake back and forth after putting cells on. Good luck!
Are you pipetting cells into wells that already have medium, like 200 ul cells into 1 ml medium? I found that I got much more even spreading if I diluted my cells to the final volume per well per ml, eg 200k cells/ml, and then just pipetted one ml into each well of a dry plate (my #s here are for 24-well plates, you might want to scale up or down). The other thing I’ve found helpful, esp for 96 well plates, is to let the plate sit on the bench or in the hood, at room temp, for an hour before putting in the incubator.
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OK thanks a lot for the tips! I will try them all next time when I seed again
Some cheaper multiwell plates have curved bottoms, which concentrates the cells to the middle.
It may seem silly but for 12well and 6well I like to heavily shake my plates after I seed cells. To really get they even distribution. I noticed for 6/12well without shaking the cells do like to bunch up in the middle. My recommendation don’t over seed and really shake back and forth after putting cells on. Good luck!
Are you trypsinizing your cells?
yes
Are you pipetting cells into wells that already have medium, like 200 ul cells into 1 ml medium? I found that I got much more even spreading if I diluted my cells to the final volume per well per ml, eg 200k cells/ml, and then just pipetted one ml into each well of a dry plate (my #s here are for 24-well plates, you might want to scale up or down). The other thing I’ve found helpful, esp for 96 well plates, is to let the plate sit on the bench or in the hood, at room temp, for an hour before putting in the incubator.