adding drug to 6 well plate
Posted 30 March 2013 - 05:18 AM
Posted 30 March 2013 - 08:16 AM
I need to add drug to my 6 well plate with cells ( each well can hold up to 3.2 ml). the molecular weight of the drug is 357 and i am given 5 mg of the drug that needs to be dissolved in DMSO. plate 1 is control, plate 2 needs to have .001 micromolar of the drug, plate 3 .01 micromolar, plate 4 .1 micromolar of the drug, plate 5 1 micromolar and, plate 6 5 micromolar of the drug. I know that i need to come up with a reasonable volume of DMSO to dissolve the drug in and that i need to start with the highest concentration (plate 6) and from there i need to make serial dilutions to obtain correct drug conc. that needs to be added to the corresponding well. And the volume of DMSO that will be added to each well has to be equal. I need to have a step by step tutorial on how to solve this problem. Thanks for help in advance!!!!
One thing you could do is a pilot study where you just test for the toxicity effect of your vehicle (DMSO). Just take your cells and add different volumes of DMSO and then count them after incubating for 2 days to get an idea of toxicity. It should give you an idea of what types of volume of DMSO you can add to your cells and avoid significant toxicity from the DMSO. DMSO is definitely toxic. When we have an extremely lipophilic molecule that needs DMSO, it is typical for us to create 100 mM stock solutions and add the volume needed to reach the desired drug concentration.
You're working on the nM range so take into consideration the fact that you want to add a volume that won't be too small (i.e. the volumes require aren't say 0.01 uL). There's simply too much error and uncertainty when working with small volumes when using a typical hand pipette.
Edited by Ubiquitous, 30 March 2013 - 08:18 AM.