Hi everyone, I´m working (at least i´m trying to) on the vaccination model for turkeys and I need to make dilution to get 200000 cells/ 200microliters of medium. If I have counted, for example, 6750000 cells/10ml of medium how do I get the number from above? This is probably very simple but I usually dont work with cells. Thanks!
1ml=1000ul
10ml=10,000ul
so, you have 6750000 cells/10,000ul
6750000 present in = 10,000ul
200000 present in= 10,000 x 20,0000 / 6750000
you will get =296.29 ul
means your required number of cells 200000 are present in 296.29ul
Take this volume,centrifuge it and add 200 ul medium.
More easiest way is that initially you should not take cells in 10 ml,rather take cells in 5ml and dilute when it is needed after counting and you do not need to centrifuge it again.
U need 100 00 00 cells /mL (200000x5).
675 00 00/100 00 00 = 6.75mL
Resuspend the cells in 6.75mL medium
Thsn u have it at 1000000/mL or 200000/200µL concn.
1ml=1000ul
10ml=10,000ul
so, you have 6750000 cells/10,000ul
6750000 present in = 10,000ul
200000 present in= 10,000 x 20,0000 / 6750000
you will get =296.29 ul
means your required number of cells 200000 are present in 296.29ul
Take this volume,centrifuge it and add 200 ul medium.
More easiest way is that initially you should not take cells in 10 ml,rather take cells in 5ml and dilute when it is needed after counting and you do not need to centrifuge it again.
Thank you very much Manoj and Sumreen.
@Sumreen - the practice of the clinic is that we grow our cultures in 10mL of medium. When performing passage, we take 1mL of culture in medium and passage it to fresh 9mL. Anyway, both ways you guys described are gonna be useful.
this the best way to make dilutions... no confusion or need of calc...
No mate, I am not offended by any means. It´s just I have no experience in lab work, during my degree I was concentrated on field practice, unfortunately. Ye i know it´s simple mathematics but at the moment I am like Alice in Wonderland and everyone has his mind blocked from time to time:)
I second the C1V1=C2V2 (though I learned it slightly differently); it is the most-often used formula in my labs. It will work with any concentration (mg/mL, %, M, ppm, etc) and/or any volume (mL, L, gal, etc.) as long as you have the SAME UNITS on BOTH sides.
Heyy Tarik...I agree with Sumreen...See the suspension that you have (6750000/10.0 ml) is more dilute than the required no. of cells (200000/200 miroL or 10000000/10.0 mL) so you will have to centrifuge your suspension and resuspend it in fresh medium..
hey tarik .......i agree with sumreen and viraj they both did good job for clearing all the concept and logic behind dilutions but i would like to share one thing, Dear in dilution be aware about the concentration of both solutions. here in your case your stock solution is dilute having 675 cell/ul (6750000 cell/10ml) and you want to prepare concetrate solution should have 1000cell/ul (200000cell/200ul). practicaly it not possible so you have to fallow calculation done by sumreen and viraj ok.
I wrote in very simple way so that anyone can understand the calculation easily whether you need it for passage or any assay like cytotoxicity assay in which you need to know the cell viability before taking a particular number of cells.Thank you Viraj and Pankaj
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