Confocal image analysis_before generating a colocalized image for analysis

Hello everyone,

I am new in the field of confocal image analysis and I really need your help in the very first steps that I have to do to normalize my images (red and green) based on my control signal (blue) and to do the threshold to my images in order to discard background noise and see my signal as sharper. How I start?Is there any sequence of steps that I have to do with my raw data before checking their yellow colocalization?

Thank you very much for your time :slight_smile: