The present article describes the creation and immobilization of Polyethylene imine (PEI) capsules on a glass surface. The synthesis and deposition were accomplished by short-time (3 min) one-step reaction. The preparation and immobilization of PEI spheres, was carried out using an environmental friendly method, the ultrasonic emulsification. The ultrasonic technique enables to control size and fulfillment of the internal part of the immobilized PEI spheres. Moreover, the ultrasonic emulsification method showed 100% efficiency in PEI spheres creation, which means no residues of aqueous PEI and oil solvents remained in the reaction flask after the nanosphere's creation. The immobilized PEI spheres have sizes varied from 50 to 500 nm. The PEI spheres were successfully filled either with organic solvent (hydrophobic) or with water (hydrophilic). This method provides us the perspective for future encapsulation of varies molecules which have hydrophobic or hydrophilic nature.