How to set up firefly bioluminescence experiment?

[yueguan]

Hey,

I was wondering if you can help me with setting up an experiment with regard to bioluminescence especially with the variables/factors involved in it.

I currently have several mg/ml of D-luciferin, luciferase (firefly), ATP solution. Furthermore I also got some MgSO4, but I don't know if I am going to need it or not. If I do need it and you think I can better replace it with another metal ions, please just tell me.

The experiment will be done (preferably several times) in a cuvette of 2,5 mL because I got to work with small amounts since the costs are pretty high.

The problem is that I don't know how I should begin, because I am struggling with a lot of factors like the ratio's of the chemicals. How much D-luciferin, luciferase, how much solvent (=water at the moment) do I need, how much ATP solution do I need to add.
I don't know whether I should dilute the solution or not.
Also, the optimal pH unknown as well and I dont know how to change pH without just random guessing how much of an acid/base I should add to get to the optimal pH.

At last IF the experiment works, the cuvette will be placed in a spectrophotometer with a range up till above a wavelength of somewhere above 1000 nm, but I have genuinely no idea what the proper way of calculating the ratio's, pH, whether I need to dilute it etc.

If you can guide me through it or give me tips about how I can deal with the problems, I would greatly appreciate it.

[MadelineB]

Hello,
If you are a student in grades K-12 doing a hands-on science project, then please provide more detail about your research plan including your hypotheses and the experiments you plan.

If you are not a student, then there are other places on the internet where your questions would be more appropriate. The experts here help students, however you are welcome to browse the Science Buddies projects, including this one:

https://www.sciencebuddies.org/science- ... background

You might also study the Science Buddies project guide:
https://www.sciencebuddies.org/science- ... ience-fair

Thank you.