Microemulsions are increasingly being used as methods of preparation of nanoparticles, especially nanospheres, due to their properties such as thermodynamic stability, optical transparency and low viscosity. Microspheres are spherical particles ranging in size from 1 to 1000 μm, characterized by a large specific surface area and biodegradability, which makes them suitable drug carriers for controlled release of drugs in smaller dosages. Microspheres can be prepared by various methods, and this work describes the preparation of chitosan-based microspheres using the emulsion, specifically the microemulsion method. A solution of chitosan, a natural polymer, was used as aqueous (dispersed) phase with the addition of copper (II) ions (Cu2+) as a physical crosslinker (solution of the CHT‒Cu complex). Isooctane and a mixture of viscous and highly liquid paraffin were used as the oil (continuous) phase, with the addition of the SPAN 85 as an emulsifier. The influence of different oil phases, the water-to-oil phase ratio and type of a reactor were examined on the composition, appearance, morphology and size of the particles. FTIR analysis showed that the obtained particles are chitosan-based, and the absence of new absorption bands indicated electrostatic nature of the interactions between chitosans’ functional groups and copper. SEM analysis showed that the obtained particles are non-porous and have smooth surface when prepared using the flat bottom reactor, with the ratio of water-to-oil phase of 1/5 and isooctane as the oil phase. Also, that system showed the narrowest particle size distribution. The particles formed in the emulsion with a mixture of viscous and highly liquid paraffin as the oil phase were irregular in shape.