The process of obtaining alginate microspheres (AMs) by emulsification method was optimized by applying statistical analysis software. Ten batches of microspheres were prepared using the fractional plan 3 ^(K-p). AMs were obtained with two different methods: an ultrasonic homogenization (UH) process and a rotor-stator mechanical homogenization (MH). The amount of a cross-linking agent (CaCl ₂), calcium chloride rate addition, and the sonication amplitude (UH) or the speed of rotor rotation (MH) were selected as formulation variables. All the batches were evaluated in terms of stability and size of the alginate microspheres. Approximation profiles were developed. As a result of the conducted research, stable alginate microspheres with sizes ranging from 10 to 30 micrometres were obtained. The obtained results showed that the quality of AMs was mainly affected by the concentration and the rate of calcium chloride addition into the system. Therefore, the role of calcium ions in the mechanisms of shell structuring was discussed. Lactobacillus casei bacteria were encapsulated into the batches found to be optimum. The high encapsulation efficiency (EE) of the bacteria (72-94%) depending on the form) and their viability over time were obtained. The model developed in the study can be effectively utilized to achieve the AMs formulations.