Resonance Raman spectroscopy (RRS) and reflection spectroscopy (RS) are optical methods applicable to the non-invasive detection of carotenoids in human skin. RRS is the older, more thoroughly validated method, whereas RS is newer and has several advantages. Since collective skin carotenoid levels serve as a biomarker for vegetable and fruit intake, both methods hold promise as convenient screening tools for assessment of dietary interventions and correlations between skin carotenoids and health and disease outcomes. In this manuscript, we describe the most recent optimized device configurations and compare their use in various clinical and field settings. Both RRS and RS devices yield a wide range of skin carotenoid levels between subjects, which is a critical feature for a biomarker. Repeatability of the methods is 3-15% depending on the subject's skin carotenoid level and the uniformity of its local distribution. For 54 subjects recruited from an ophthalmology clinic, we first checked the validity of the relatively novel RS methodology via biochemical serum carotenoid measurements, the latter carried out with high performance liquid chromatography (HPLC). A high correlation between RS skin and serum HPLC carotenoid levels was established (RâŻ=âŻ0.81; p < 0.001). Also, a high correlation was found between RS and RRS skin levels (RâŻ=âŻ0.94 p < 0.001). Subsequent comparisons of skin carotenoid measurements in diverse age groups and ethnicities included 569 Japanese adults, 947 children with ages 2-5 screened in 24 day care centers in San Francisco, and 49 predominantly Hispanic adults screened at an outdoor health fair event. Depending on the particular subject group, correlation coefficients between the RRS and RS methods ranged between R âŒ0.80 and R âŒ0.96. Analysis of the Japanese screening showed that, on average, skin carotenoid levels are higher in women compared to men, skin levels do not depend on age, and tobacco smokers have reduced levels versus non-smokers. For the two most ethnically diverse groups with widely varying melanin levels, we investigated the effect of dermal melanin on RS and RRS skin carotenoid levels. The analysis revealed that large variations in skin carotenoid levels remain detectable independent of the particular melanin index. This behavior is consistent with the absence of melanin effects on the skin carotenoid levels generated with the instrument configurations. The RS method has an advantage over RRS in its relative simplicity. Due to its detection of skin reflection over a wide spectral range from the near UV to the near IR, it has the unique ability to quantify each of the major tissue chromophores and take them into account in the derivation of skin carotenoid levels.