I want to perform an MTT assay for a cell proliferation study. After culturing the cells, when we need to seed the cells in a 96 well plate then how would I calculate the volume of cell containing medium needed for that?
Simply after you count the cells you just calculate the volume of cells needed for 96(100 microliter per well) so ( volume of the needed cells =the desired volum .the desired number of cells / the orginal counted cells /ml ) in this way you gonna have the nummer of cells needed for 96 well plate ...good luck
Thank you very much. Actually i want to seed 3000 cell/well. But i want to know the amount of volume of medium (with cultured cells) required for that.
Firstly you have to count the cells cultured in the flask and then calculate the concentration; then you have to calculate the volume of cells needed (for example if you seed the cells in 96 wells you need 100 μL∙96 wells=9600 μL of cells suspension).
I usually prepare the cells suspension in a Falcon tube in this way:
- I calculate the volume of orginal counted cells needed for the seeding→ = (final concentration of cells ∙ total volume of cells suspension needed) / concentration of original counted cells → in your case the final concentration of cells is 30000cells/ml and the total volume of cells needed is 9600 μL (if you seed 96 wells)
- I prepare in a Falcon tube the medium (total volume of cells suspension needed– volume of orginal counted cells needed for the seeding) and add the calculated volume of original counted cells needed (gently mixing to have homogeneous suspension)
- I put 100 μL of this suspension in each well
I believe you are interested in seeding 3000 cells/well.
In my opinion, first we need to know the total cell harvsted via trypsinization.
Then, we would know the cell concentration as highlighted by @Marta Serena.
Let say the cell concentration is 100 cells/uL.
Next, we would want to know the number of cells needed for your experiement.
Let say you would want to seed 3000 cells in 10 wells, so this means that we need a total of 30000 cells.
Then, in order to know the volume needed to take from cell suspension to supply 30000 cells, we should divide 30000 with 100. So, 30000/100 = 300 uL.
From cell suspension, 300uL is needed to supply us with 30000 cells. Meaning each well will receive 30uL.
As our colleagues have described, the recommended volume for each well should be 100uL. So, after putting 30uL in each well, the remaining 70uL can be the culture media.
I hope this helps, and do excuse me if I bore you too much.
All the best. Thanks. =)
Thank you very much for the ans making it easy for me. wish you all the best.
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