I've already run micro dilution MIC test using 96 wells and some of the wells gave turbidity for a particular antimicrobial plant extract, but I can't know whether it is given by live cells or insolubility of antimicrobial compound in MHB.What are the possible tests that I have to conduct? Thank You in advance.
You can try with LB-Agar (0.15%). If your compound does not solubilize, you can use DMSO but you have to take into account that the final volume of DMSO does not exceed 1%.
Thanks you very much for your suggestions (Stefan Zimmermann
Abdelhakim Bouyahya)I''ll try with DMSO
It depends on the tested compound. You can use DMSO, Methanol, ethanol ... at a very low final concentration (< 2%), with a negative control.
Or you can determine the MIC by the agar dilution method.
First prepare your stock solution 100 times higher than the working concentration with absolute DMSO, then diluted 100 times with your medium. If the compound still turbid run along controls made of serial dilution of your compound with only medium( without microorganis) and cancel the absorbance of the control from the tested one. OR use agar dilution method instead of broth dilutlion method to find the MIC.
I used to solubilize the plant extract in DMSO (2 %). You can try it and don't forget to test whether the DMSO at this concentration will be toxic to your microorganisms.
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