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How to achieve uniform cell distribution in culture vessle

Cell Culture Colony Formation Assay seeding

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5 replies to this topic

#1 shamash119

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Posted 13 November 2012 - 09:29 AM

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!

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#2 Baraa

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Posted 13 November 2012 - 09:34 AM

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

#3 bob1

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Posted 13 November 2012 - 11:49 AM

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

#4 leelee

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Posted 13 November 2012 - 06:39 PM

I agree. Don't swirl the plate.

#5 pcrman

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Posted 13 November 2012 - 10:32 PM

There is a long thread on this topic here http://www.protocol-...-6-wells-plate/

#6 shamash119

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Posted 14 November 2012 - 08:59 AM

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?

Edited by shamash119, 14 November 2012 - 09:12 AM.






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