Understanding the molecular mechanisms underlying bubble-(bio)surfaces interactions is currently a challenge that if overcame, would allow to understand and control the various processes in which they are involved. Atomic force microscopy is a valuable tool to measure such interactions, but it is limited by the large size and instability of bubbles that can be attached on surfaces or on AFM cantilevers. To overcome these challenges, we here develop a new method to probe more accurately the interactions between bubbles and (bio)-interfaces by taking advantage of the fluidic force microscopy technology (FluidFM) that combines AFM with microfluidics. In this system, a micro-sized channel is integrated into an AFM cantilever and connected to a pressure controller system, thus creating a continuous and closed fluidic conduit that can be filled with a solution, while the tool can be immersed in a liquid environment [1]. An aperture at the end of the cantilever allows liquids to be dispensed locally. In this study, we use FluidFM in an original manner, to produce microsized bubbles of 8 µm in diameter, directly at the aperture of the microchanneled FluidFM cantilevers. For that, as shown in Figure 1 instead of liquid, the cantilever is filled with air and immersed in a liquid environment. By applying a positive pressure inside the cantilever, we succeeded in forming bubbles of controlled size directly at its aperture. Because the same pressure is maintained in the cantilever during the experiment, the dissolution of the gases from the bubble is compensated, which allows keeping the size of the bubble constant over time. After the characterization of the bubbles produced using this method, their interactions with hydrophobic surfaces were probed, showing that bubbles behave like hydrophobic surfaces. Thus they can be used to measure the hydrophobic properties of microorganisms’ surfaces, but in this case the interactions are also influenced by electrostatic forces. Finally we developed a strategy to functionalize their surface, thereby modulating their interactions with microorganisms’ surfaces. This new method provides a valuable tool to understand bubble-(bio)surfaces interactions but also to engineer them.