Synapses represent a unique neuronal compartment specialized in communication. The morpho-functional investigation of the synapse has always been hindered by its tiny size and high density of molecular elements, but specific technological and methodological advances helped us to reduce these limitations. As the matter of fact, in recent years, we have witnessed the continuous development of new methods allowing measuring and controlling synaptic activation both in vitro and in vivo. These include new genetically encoded sensors of synaptic exo-endocytosis, but also engineered synaptic proteins able to inhibit the release of neurotransmitters. Furthermore, new promising tools allow changing the functional properties of synapses in a plasticity-like manner. All these methodological advances are likely to generate unprecedented knowledge about the dynamics of synaptic transmission and plasticity at several levels of the nervous system. Nevertheless, in most cases, the implementation of these new methods remains technically demanding, likely due to the high complexity of their operating principle. Hence, it would be worth refining and potentiating these tools in order to extend the range of experimental settings for their application.