dilution of cell suspension - (Oct/07/2012 )

hi
I want to seed a 96 well plate from a T25 flask having a volume of 4mL. Each well should have 10,000 cells in 100uL. I trypsinised the cells and added 1mL of medium to get a cell suspension. Its concentration is 1e6 cells/mL. So I took the entire 1mL of cell suspension and diluted it with 9mL of medium. From this I added 100uL into each well so that it contains 10,000 cells in 100 uL. Is this correct?

10⁶ cells in 10 ml will be 10⁵ cells/ml = 10⁴ per 100 ul. therefore correct.

thanks for the reply bob. One more doubt. the concentration which I got (1e6 cells/mL) is the concentration of the 1mL cell suspension right, not of the initial 4mL in T25 flask right?

The concentration of the cells is measured in the volume you have collected them in after you trypsinized them, not on the volume of medium you grew them in.

thanks bob. that helps me a lot.