I have a dried lyophilized sap from a plant called Croton lechleri Mull. Arg.
I want to treat cells in different concentrations (groups): 0.01, 1, 2.5, 5, 50, 100 ug/mL. However, I am not sure how to translate that into the cells. I intend to use a 96 cell plate with 100 uL/well.
Cells lines: BV2
DMSO purity 99.99%
Croton lechleri solubility in DMSO: 10 mg/ml
How much do I need for my stock solution (so I can treat the cells in these concentrations)? Can someone please explain how this is calculated? Thank you
just try a low concentration and a high dose, do a pertest before your real test
I like Miao Zheng's idea, but with one change. I recommend you pre-test with only a negative control (zero sap). You want to observe any effects on your cell line produced by the highest concentration of DMSO you will test. To get that number divide your highest sap concentration sample by your stock concentration: 100 ug/mL / 10 mg/mL = 0.01, or 1%. So I recommend you start by testing 100 ul of a 1% solution of DMSO (diluted in your growth or harvest medium) against your cells. DMSO generally is not well-tolerated by cell cultures as it dissolves cell membranes, but you may be able to use it in a brief assay. You need to know about any changes to your cells. So check the DMSO-exposed cells in your assay, but also look at them under the microscope. If they seem "normal," we can set up a simple dilution series for your test samples.
Let me warn you that making dilutions of DMSO into water is tricky, because DMSO dissolves in water (about 70%). If you do end up using DMSO as your carrier solvent I recommend you prove this awkward fact to yourself by mixing 10 mL of DMSO to 10 mL of water and then measuring the final volume in a graduated cylinder. It is not 20 mL, as you may expect; it's closer to 13 mL. So if you use DMSO for your stock solution you'll need to account for this phenomenon when you make your dilutions. For samples at low concentrations the effect will be insignificant, but for higher concentrations you will want to estimate the effect to avoid a large systematic error.
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