Cell plating calculation !! - Help needed with Cell plating calculation !! (May/05/2009 )

Hello,

I need help with cell plating calculations.

I have to plate 5 million cells per well. Each well with 3ml. Altogether I need 6 wells.

Currently, I have 200ml cells with 5.2 x 10^5 cells/ml.

How to plate my cells? Do I have to do dilutions?

Thanks,

Ok, so you need 5 million cells per well in 3ml.

So, you have 5.2 x 10^5 cells per ml.
Divide 5 million cells (5 x 10^6) by 5.2 x 10^5 and you get 9.61- which means you would need to add 9.61 mls of your cell suspension to each well to get your 5 million cells.

I recommend pelleting your cells, then resuspending in a much smaller volume.

You have a total of 1x 10^8 cells (5.2x10^5 multiplied by 200).
I would resuspend in approx 20ml (1x10^8 cells divided by 5x10^6 cells= 20, so resuspend in 20mls for cells at 5x10^6/ml)

So add 1ml of your new suspension to each well and top up to 3ml (or you could resuspend in 60ml and aliquot out 3ml of your suspension per well)

I hope this makes sense

(and I also hope I haven't made any mistakes!!!)

C=N/V and C_iV_i=C_fV_f are your friends for these calculations. C= concentration, V=volume, N= total number, i=initial, f=final.