Carotenoids are a group of natural pigments, consisting of more than 750 compounds. They are mostly yellow, orange, or red in color, due to the system of conjugated double bonds. This structural element is also responsible for the good antioxidant properties of many carotenoids. Carotenoids have shown numerous biological activities (not only as provitamin A), e.g., preventive properties of fruits and vegetables. As lipophilic compounds, their uptake and storage in the body are dependent on various conditions. In vitro and in vivo data showed stimulating and inhibitory effects of matrix compounds on bioaccessibility and bioavailability of carotenoids. This Special Issue presents the most recent advances in carotenoids research, in addition to the search for antioxidant properties. Chapters present the photoprotective properties of carotenoids as well as the activities of carotenoids related to liver health. Research data on the effect of degree of ripeness on carotenoids pattern in rosehip and possibilities to use shrimp waste as source of carotenoids are presented. Other investigations characterized apocarotenoids in microalgae and the properties of inclusion complexes of lycopene and beta-cyclodextrin. Biological activities of synthesized retinoyl-flavonolignan hybrids were also reported. In addition, the effects of in vitro digestion of human milk on the micellization of carotenoids were investigated.