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#1 bio08

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Posted 15 April 2010 - 01:48 PM

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!

#2 bob1

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Posted 15 April 2010 - 04:46 PM

#3 rkay447

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Posted 15 April 2010 - 08:06 PM

bio08, on Apr 15 2010, 03:48 PM, said:

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!

The way you are phrasing your question is very confusing.  So, to start, do you have TWO cell samples that you need to dilute to 1 mil/mL??  I bolded the confusing part of your post.  You say that you have a cell pellet that you
resuspend in 5mL media and it is 250 mil/mL but then your next line says that you counted the cells and have 1.8 mil/mL.  It's not clear what your samples really are and what you are trying to do but I'm going to try to help.  You want a final cell suspension of 1 million cell/mL, right?  And, you need 10mL of this 1 mil/mL, so you need 10 million cells.  If your cell suspension is 250 mil/ml you need to figure out what volume will equal 10 million.  Divide 250 million by 1000 tells you how many cells you have per uL.  This is 250,000 cells/uL.  Divide 10 million by 250,000 and you get 40.  You need 40uL of the 250mil/mL cell suspension to get 10 million cells.  Add this to 9960uL media and you have a 1 mil/mL suspension.  Now, if you counted your cell suspension and saw that you actually only have 1.8 mil/mL this means you only have a total of 9 million cells in the 5 mL and you are not going to be able to make 10mL of a 1 mil/mL sample.  Sorry, not enough cells.  I hope this helped and didn't confuse you more!