cell count - (Jun/10/2011 )

I have counted cells on hemocyt. It is 5x10^5 cells. I need to plate them at 2X10^6 per well in 1 ml of media in 96 well plates. How many microlitre of the cells I need to add to the 1 ml of media on the plate?

Assuming that your cell suspension has 5x10^5 cells per ml.., and the required conc is 2x10^6 in the final volume 1 ml, you need to pellet down and resuspend it in a lower volume say about 10:1 and from that you might need to add 400 micro litres...

Gnana..

Limpa on Fri Jun 10 12:01:47 2011 said:

Okay so your question doesnt really make much sense, there are some issues with it. I'll try to go in steps and see if we can get somewhere.

You say you have 5x10^5 cells.... is that in total?

If it is in total: you dont have enough cells to make them 2x10^6cells

Further to that, you say you need 2x10^6 cells per well in 1ml in 96 well plate. Now, do you mean 2x10^6cells/ml... or 2x10^6cell/well. because there's no way you can plate 1ml in a well of a 96 well plate, usually you plate 100-200ul tops.

Assuming you plate 100ul per well, you will need ~10ml per plate.

IF what you actually need is 2x10^6 cells/ml you need to have 2x10^7 cells in total. Which, back to the beginning you dont have, as you say you have 5x10^5 cells.

IF what you need is 2x10^6 cells/well you will need 2x10^6 x96 = 192x10^6 ~ 2x10^8 cells. Once again, not enough if you have 5x10^5 cells.

Now, if you actually have 5x10^5cell/ml you will need to spin down your cells and resuspend them again into the appropriate volume for the concentrations I said above. For this use C1V1=C2V2 where C1=5x10^5 V1=the volume of cells you have C2=the concentration you need and V2=the volume needed for the 96 wells.

I hope this helps.

Thanks guys