Carotenoids are essential pigments in plants that are primarily stored in plastids, mostly chloroplasts and chromoplasts. Understanding the ultrastructure of plastids, particularly chromoplasts, is of importance when assessing carotenoid bioavailability. Chromoplasts, which originate from precursor plastids like chloroplasts, proplastids, or amyloplasts, undergo structural changes during maturation, which includes carotenoid accumulation and membrane remodeling, varying among plant species and tissues. Chromoplasts are classified according to their ultrastructure in globular, tubular, and membranous types. The deposition form of carotenoids within chromoplast structures, such as plastoglobules and crystals, influences their bioavailability and stability. Crystals formed by carotenoids, such as β-carotene and lycopeneLycopene, may be observable via light microscopy. On the other hand, tubular, globular and membranous chromoplasts require transmission electron microscopy (TEM) for accurate characterization due to their fine ultrastructure. Light microscopy is a rapid method that may be used to assess carotenoids in plant tissues without staining. Sample preparation consists of simple techniques like hand sectioning or squashing. On the other hand, TEM gives a sub-Ångström resolution for ultrastructural analysis, and needs specialized sample preparation including fixation, embedding, sectioning, and staining. TEM analysis requires imaging-stained samples at appropriate magnifications and voltage settings, followed by image analysis and interpretation. Recent developments in TEM technology permits large-scale datasets facilitating exploration of carotenoid deposition in plastids. Combining light microscopy and TEM techniques provides better insights of carotenoid deposition in plastids.