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Question about more complicated splittings - (Jun/26/2012 )

I'm trying to figure out a methodology for splitting cells into multiple plates and flasks.

Lets say I have a T25 flask and two wells in a six well plate that I want to split 1:3 into a whole six well plate and two T25 flasks. My boss was adding up all the surface area or something and now I'm all confused.

How would I go about doing this?

I understand splitting ratios are just based on your initial amount of cells, but I'm getting lost in the complications.

I've tried googling and searching and looking at Freshney but nothing helps if i'm trying to split into many vessels.


A 1:3 split is a 1:3 split of the total amount always - not the area - so take your flask and the plates, lift them all and pool the cells. Spin down and re-suspend in a suitable volume. Take the final volume and split 1:3 into the vessels you have.

Note that this doesnt work for changing flasks etc. You will be much better off counting the cells and seeding at a defined density for the 6 well plates.