hi everyone,

I need your help

my cells have GFP ( transfect the cell with GFP plasmid) now I want to measure the GFP intensity along with the pH change

for example when pH changes from 5 to 6 I am assuming I will see a weak intensity peak while if It changes from 6 to 7 I am assuming the intensity peak will be strong

it is hard because 1) I can not use a buffer with a different pH that will change the outside pH not the inside so there will be no effect on my GFP intensity because the GFP is attached inside of one of the ion channels

2) I can't use a fluorescent pH indicator because the intensity I will get will be for the indicator, not GFP

bascally, I want to test if my plasmid that has GFP can sense the pH change ?

I will trigger the membrane potential then I will measere the GFP intistiy but I need a way to measre both GFP intensity and pH change.

thank you