For my project, I need to expose zebrafish larvae in petri dish to three concentrations of rotenone (10nM, 30nM and 50nM), and the solutions should be changed daily . My current method was first, preparing 1mM of rotenone stock solution from rotenone powder and 100% DMSO, then store it at -20°C. Then I do 10 fold serial dilutions from stock solution (diluent: E3 media) until I reach 1000nM and prepare the 3 concentrations. I repeat the serial dilutions and preparation of the concentrations daily until 7dpf. I'm thinking of making a large volume of the working solution (1000nM) and store them (maybe at 4°C?) so it's easier to prepare the solutions and to do reduce errors in dilutions, but are they acceptable?
Hey Liyana,
we use a 50 mM Stock of Rotenone in DMSO. We made aliquots with 100 µl per vial and stored them at -20°C. This stock now lasts 7 years and works very well. I forgor one of these aliquots for two months in the fridge (so at 4°C) and it still worked. We treated ex vivo macrophages with a f.c. of 100 µM, so I am not sure if it will work with your concentrations and in zebrafish, but theoretically it should.
All the best and a happy new year,
Marc
@marcherb Thank you for replying! The aliquots you mentioned, they were rotenone in DMSO right? I've been freezing the stock solution too. It's just that since the concentrations I need to use are quite small, I couldn't find a better way to prepare them.
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