Please help me to understand- cell counting - (Apr/15/2010 )

Hello,
I am a beginner so please help me to understand following:

I centifuge my cells and dissolve the pellet in 5 ml media.

In 5ml cell suspension I have 250 mil cells/ml
In this 5 ml cell suspension I counted the cells and I have 1,8 mil cells/ml

I want 1 mil/ml and want to make 10 ml cell suspension

So I have 250 mil cells in 1ml
10 ml *10^6 cells= 10 mil cells ( in 10 ml I have 10 mil cells)

10 mil cells/ 250 mil cells = 0,04 ml= 40 ul. So I take 40 ul from my 5 ml cell suspesnion, but I need 10 ml in total volume so to the 40 ul cells ( I took from my 5ml cell suspension) I will add 9,96 ml media.

Is this calcultation right or this one below:

In 5ml cell suspension I have: 5ml *250 mil cells/ml= 1250 mil cells

1250 mil/10 mil = 8 ul I will take from the 5 ml cell suspension and I will add around 9,92 ml media

Thank you!

you have 250mil cells per ml.

and you need 1 mil cells per ml, so you need to dilute it 250 times.

This means you add 249 ml to 1 ml of your start solution.
and there you go. You have 250 ml of a 1mil cells per ml solution.

If this 249ml is too much you can start with 0.5ml of the start solution and add 249/2 ml or you take 0.1ml and add 24.9ml....

It all depends on whats the easiest way for you to work.




What you are doing, I do not get it. If you take 40�l from the 250mil cells per ml then you have indeed 10mil cells ; but you have 10mil cells in your 40�l ... so if you add new volume to that 40�l you will dilute your cells and not have 10mil cells per ml. Or do you just need 10mil cells?