Intravital imaging allow us to watch cellular and molecular behaviour in higher fidelity in vivo models that more closely recapitulates the native environment of the organ or disease of interest in real-time. In this talk I will present multiple ways in which we have used Intravital imaging, proteomics, biomechanical assessments and biosensors to fine-tuned drug treatments and optimise outcomes. I will also discuss the benefits of using longitudinal imaging using titanium optical windows approaches in pancreatic cancer to understand and target tumour-TME feedback in this deadly disease.