One of the fundamental pillars of a properly set up marketing communication is customer orientation, defining their needs and preferences and then creating their profiles. Following this fact, the aim of the paper is to create subsegments of Generation Y (Millennials) in the Czech market based on identified factors in terms of lifestyle as the non-traditional segmentation descriptive variable. In order to achieve the aim of the paper, the AIO parameters within psychographic segmentation were used to process the primary data collected by the marketing research carried out in the form of a questionnaire. Overall 999 respondents who are residents of the Czech Republic participated in this marketing research. The characteristics of Generation Y (Millennials) and the determination of the main points for external and internal marketing strategies create the basis for the comparison of subsegments based on the criteria determining the possible success of the company in this process.