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dilution of cell suspension


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4 replies to this topic

#1 vennela

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Posted 07 October 2012 - 05:52 AM

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?

#2 bob1

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Posted 07 October 2012 - 12:29 PM

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

#3 vennela

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Posted 09 October 2012 - 08:34 AM

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?

#4 bob1

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Posted 09 October 2012 - 12:22 PM

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.

#5 vennela

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Posted 12 October 2012 - 09:23 PM

thanks bob. that helps me a lot.




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