To understand biological processes at the cellular level, a general approach is to alter the cells’ environment and to study their chemical responses. Herein, we present the implementation of an electrochemical push-pull probe, which combines a microfluidic system with a microelectrode, as a tool for locally altering the microenvironment of few adherent living cells by working in two different perturbation modes, namely electrochemical (i.e. electrochemical generation of a chemical effector compound) and microfluidic (i.e. infusion of a chemical effector compound from the pushing microchannel, while aspirating it through the pulling channel thereby focusing the flow between the channels). The effect of several parameters such as flow rate, working distance and probe inclination angle on the affected area of adherently growing cells was investigated both theoretically and experimentally. As a proof of concept, localized fluorescent labeling and pH changes were purposely introduced to validate the probe as a tool for studying adherent cancer cells through the control over the chemical composition of the extracellular space with high spatiotemporal resolution. A very good agreement between experimental and simulated results showed for instance, that the electrochemical perturbation mode enables to affect precisely only few living cells localized in a high-density cell culture.