I´m kind of stuck at my image analysis. I want to quantify the colocalization between two signals in different mutant Backgrounds to see if there is any change. Problems are the pretty small size of the vesicles and the noise because of the weak signal from one of the proteins. I can´t enhance the expression to get a better signal because I have to use the natural promotor. I tried with JaCoP but I get values around 10 % colocalization in wildtype when from my optical impression it has to be around 80-90 %. I also tried to reduce the noise before the calculation which brings me to 20 % colocalization. Has anybody an idea how to solve this? Some pictures are attached
Thanks a lot!