I would like to test a concentration of a graphene oxide based chemical solution for bacterial viability. I wanted to know how to conduct an experiment in agar to test the best concentration of the solution that favors the growth of bacterial colonies. It would be a test with various concentrations of the chemical solution. Thanks
Dear Carolina Brom Aki de Oliveira , This is an interesting question as normally scientists test the efficiency of antibacterial compounds. But I think you can proceed using their approach but slightly adapted.
Here's how I would do this:
1. Prepare a set of petri dishes by marking different zones on the bottom using a marker pen. These sections will contain your different dilutions so you need to be able to see how much was added to each section.
2. Next, prepare an agar that is not too rich so that growth is not too fast. Preferably the agar is transparent so you can see the marked sections easily.
3. Pour the petri dishes with agar and incubated them for a few days to safeguard they are sterile.
4. Sterilize a set of paper filter rounds of let's say 10 mm diameter and keep them sterile.
5. Prepare your test-solutions, sterilize them e.g. by filtration over 0.2 micrometer filter.
6. Carefully pipette your test solutions onto the paper filter rounds allowing the liquid to be absorbed until the paper filter rounds are fully saturated.
7. Carefully transfer the paper filter rounds onto the appropriate section of the sectioned petri dishes (I would do this experiment in at least 5-fold, better maybe 10-fold (so prepare 5-10 petri dishes), as bacteria can have a mind of their own). Don't forget to include at least two blanks (paper filter rounds saturated with sterile 0.9% NaCl solution).
8. Next, pipette one or two droplets of your bacterial onto each of the paper filter rounds. Put the petri dishes in an incubator.
9. Important now is to check everyday if growth occurs, and if it does use your phone to take pictures of petri dishes.
10. Evaluate growth by looking at the diameter of the bacterial colonies on each section compared to the blank.
This procedure is lengthy, but scientifically sound. Please check this link for how testing antibacterial agents is done: https://courses.lumenlearning.com/microbiology/chapter/testing-the-effectiveness-of-antiseptics-and-disinfectants/
Kind regards, Leo
Leo Vleugels Thank you so much for collaborating with me and taking care to help me
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