# How to seed 5000/well in 200 microliters in a 96 well plate? - (Mar/17/2011 )

Hi,

Could someone help with this??? I thought to do it like this:

5000/0.2= 2.5e4 cells/ml

If my cell count was 4.5e5 cells/ml
I suppose I can use CxV=CxV
So: 4.5e5xV=2.5e4x(0.2x100)=1.1ml
So I will have to take 1.1ml of the 4.5e5 cells/ml and just add 0.2ml to each well in the 96 well plate?

Thanks!

-labt-

Hola, yes you need a 25000cels/ml suspension .And you have a 450.000 cels/ml.So you have to dilute 1/18 . So 1ml of yuor cell susp+ 17 ml of medium and you have enough volume to 90 wells at 200ul/well with 5000 cells each. Buena suerte

-protolder-

protolder on Fri Mar 18 06:33:59 2011 said:

Hola, yes you need a 25000cels/ml suspension .And you have a 450.000 cels/ml.So you have to dilute 1/18 . So 1ml of yuor cell susp+ 17 ml of medium and you have enough volume to 90 wells at 200ul/well with 5000 cells each. Buena suerte

Thanks a lot, but one more thing it should not be to dilute my cell suspension 1/20, so I should take 1ml of my cell suspension+19 ml of media so I will have enough for 96 wells?

-labt-

labt on Mon Mar 21 16:29:42 2011 said:

protolder on Fri Mar 18 06:33:59 2011 said:

Hola, yes you need a 25000cels/ml suspension .And you have a 450.000 cels/ml.So you have to dilute 1/18 . So 1ml of yuor cell susp+ 17 ml of medium and you have enough volume to 90 wells at 200ul/well with 5000 cells each. Buena suerte

Thanks a lot, but one more thing it should not be to dilute my cell suspension 1/20, so I should take 1ml of my cell suspension+19 ml of media so I will have enough for 96 wells?

Yes you colud do it, but you will have 4500cels/well that is about the same, because the numbers of cells/well are aproximatives. Buen día

-protolder-

protolder on Tue Mar 22 07:00:46 2011 said:

labt on Mon Mar 21 16:29:42 2011 said:

protolder on Fri Mar 18 06:33:59 2011 said:

Hola, yes you need a 25000cels/ml suspension .And you have a 450.000 cels/ml.So you have to dilute 1/18 . So 1ml of yuor cell susp+ 17 ml of medium and you have enough volume to 90 wells at 200ul/well with 5000 cells each. Buena suerte

Thanks a lot, but one more thing it should not be to dilute my cell suspension 1/20, so I should take 1ml of my cell suspension+19 ml of media so I will have enough for 96 wells?

Yes you colud do it, but you will have 4500cels/well that is about the same, because the numbers of cells/well are aproximatives. Buen día

Muchas gracias

-labt-