What is the best way to seed cells in 24 well plates? Can we seed in the all wells? Why people don not use the outer edge well for cell seeding?
Hi,
Normally wells are not skipped in 24-well plates but skipped in 96-well ones. We normally just put sterile water on the outside wells of 96-well plates. This is done because the volume of the medium is very little (~100ul) in the 96well plates and the wells on the outside often lose some volume due to evaporation, leading to variation in result. In the 24-well plate 400-600ul medium is used and variation is not as much as in the 96-well plate.
Hi,
Normally wells are not skipped in 24-well plates but skipped in 96-well ones. We normally just put sterile water on the outside wells of 96-well plates. This is done because the volume of the medium is very little (~100ul) in the 96well plates and the wells on the outside often lose some volume due to evaporation, leading to variation in result. In the 24-well plate 400-600ul medium is used and variation is not as much as in the 96-well plate.
I normally skip the outer cells when i am using the plates to be viewed under IVIS
I will also consider the cell type how “happy” they are at that volume, the treatment type, the period of time of incubation, or the assay type you are going to use on the plate.
When cultures are maintained over several days in a 24 well plate, the media can evaporate significantly, particularly around the edges. This can lead to all sorts of variation with treatment and/or analysis of the media from well to well. It is wise to keep sterile water in the outer edges.
What Jonathan wrote is right, but with a 24well plate too many wells are lost if we put water around the edges. I would think most of the time medium would be changed daily. Of course, see how much variation there is between wells etc, as suggested by Jonathan.
Thank you all. but for my assay my culture time will not be more then 24 hour. In this case can i use all the well. As Aniko mention we will lost many wells. If u dont use edge wells then working well will be only 8. But I have to use 4 concentration for test compound and also for triplicate.
Hi,
we do not see any real difference in cell numbers when we use all 24 wells and incubate for 24hrs. I am confident that you should be able to use all wells. If you see major difference repeatedly then consider not using those wells.
Cheers
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