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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!

-shamash119-

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

-Baraa-

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

-bob1-

I agree. Don't swirl the plate.

-leelee-

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/

-pcrman-

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?

-shamash119-