How to achieve uniform cell distribution in culture vessle - (Nov/13/2012 )

Hello all,

I am having some difficulty in achieving uniform cell distribution after seeding into 6-well plates. It seems that after swirling that the cells adhere to the edges, I have attached a picture for you to view.

Any ideas how to achieve uniform cell distribution to the center and not just the outside and stop this ring formation.

Thanks again!

After seeding your cells, do not swirl the plate. Pipet gently one or twice to obtain a homogenous cell distribution in the media.

Also note that rings or other regular patterns in your flasks/plates indicate that there is a vibration problem in your incubator.

I agree. Don't swirl the plate.

There is a long thread on this topic here http://www.protocol-online.org/forums/topic/6155-how-to-seed-the-cells-evenly-in-6-wells-plate/

Hi guys I have read in other forums that this is the best way?


Sip up whole media(wit unevenly distributed cells in the well) in to a tip n release them again 2 to 3 times. it wrked for me.for larger plates up n down left n rite rocking for 3 times is enuf.


Also -you put your cell suspension in the well,
-pipette a little bit,
-then put the lid on the plate and
-draw an "eight" with your plate as a "pencil" and the hood as a "notebook"

Also rocking back and fourth/side to side, then do the figure eight technique with the plate, and lastly let the plate sit for about 40 minutes in the hood before I proceed to move the plate into the incubator.

Does anybody have have any preferences?

Letting the plate sit outside for 40 minutes would make the cells unhealthy.