Optical imaging has revolutionized our understanding of how biological systems behave at a molecular level. In our group we develop Dynamic Activatable Fluorophores (DYNAFLUORS) as chemical probes to image molecular events associated to inflammation and cancer. With DYNAFLUORS, we aim to interrogate fundamental biology in real time as well as provide better tools to diagnose and treat disease in the clinic. We create DYNAFLUORS using a multidisciplinary approach that involves organic and peptide chemistry, fluorescence spectroscopy, cell biology and optical imaging. DYNAFLUORS are excellent imaging probes because they emit a fluorescent signal only after they interact with their target molecules (e.g., proteins, enzymes) or in specific microenvironments (e.g., organelles, high/low pH). With this approach, we obtain high signal-to-noise ratios and increased sensitivity, which allow DYNAFLUORS to be used in small doses, reducing any potential adverse effects and facilitating the translation to the clinic.