Carotenoids, precursors of vitamin A, are natural pigments that plants and some microorganisms synthesize. Thus, this chapter aims to present the current status of carotenoids and their future prospects. This study also provides a detailed analysis of carotenoid biosynthesis and its applications in domestic and industrial settings. Carotenoids are mainly used in the food and beverage, food supplement, cosmetic and pharmaceutical industries due to their numerous bioactivities. Humans and animals are not able to synthesize and need to acquire them by alimentation. Beyond the health benefits promoted by vitamin A, these compounds have antioxidant capacity, anti-inflammatory and antitumor, which protects them from cardiovascular and degenerative diseases, reduces risk of cancer, chronic inflammation, macular degeneration, and signs of aging, strengthens the immune system, and improves cognitive function. The presence of conjugated double bonds in carotenoid structure contributes to their pigmentation, absorption of ultraviolet/visible radiation, and antioxidant activity, but they are also the main reason for their chemical instability. Some factors, such as heat, light, acids, organic solvent reflux, crystal melting, and iodine treatment, can promote carotenoid cis-trans isomerization and oxidation, and therefore must be carefully considered in a process of obtaining carotenoids. In the qualitative and quantitative analysis of carotenoids, extraction and storage steps are important to prevent degradation and total recovery. The extraction is generally realized using organic solvents in addiction or not a saponification step. High-performance liquid chromatography is the most widely used technique, but others may be employed. The industrial production of carotenoids can be made by simple extraction or microbial process. In this sense, this chapter provides an overview of recent updates of carotenoids’ market, health benefits, biosynthesis, modification, analytical methods, as well as sources and production methods.