I am carrying out cytotoxicity assay on THP1 cell and I have to add a drug at varying concentration. I am wondering how can I maintain the same volume for different concentration because my drug is not really concentrated and to raise my treatment concentration to the max that I need, I have to add up to 80ul of the drug which is aqueous solution to a well of 96 well plate containing 100ul of cell and media. I found this affects the result due to the media dilution. I tried to make a double concentrate of my media using powder media so that making a 1:1 2X media with the drug will give 1X media but half the concentration of the drug. now I have to add up too 160ul of this to achieve max concentration and this also affects the result. Not sure what to do at this point. Any help is highly appreciated.
You need to use the (dry) powdered form of the drug. Weigh different amounts of the drug using the analytical balance. Then dissolve each portion in the same volume of buffer. Then add equal volumes of media+cells to each. The final volumes should all be equal or nearly so. HTH.
Thanks Dr Vicente. I really appreciate your contribution. The problem is the drug is a nanoparticle I synthesized and its in aqueous state that cannot be powdered.
Perhaps you need to try and devise a way to dry or lyophilize the nanoparticle. If this has never been done before, this is a potential publication.
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