Background: Objective biomarkers are needed to assess adherence to vegetable and fruit intervention trials. Blood carotenoids are considered the best biomarker of vegetable and fruit intake, but collecting blood is invasive and the analyses are relatively expensive for population studies. Resonance Raman spectroscopy (RRS) is an innovative method for assessing carotenoids in skin noninvasively. Objective: Our objective was to compare blood carotenoid concentrations with skin carotenoid assessments by RRS during a controlled feeding intervention. Design: Twenty-nine participants consumed low-carotenoid diets (6 wk, phases 1 and 3), a provided diet containing 6-cup equivalents (1046 g/d) of vegetables and fruit (8 wk, phase 2), and usual diet (final 8 wk, phase 4). Results: At baseline, skin and plasma total carotenoid values were correlated (r = 0.61, P , 0.001). Skin and plasma carotenoid values decreased (P , 0.001) 36% and 30%, respectively, from baseline to the end of phase 1 and then increased (P , 0.001) by .200% at the end of phase 2. Plasma carotenoids returned to baseline concentrations by the middle of phase 3 and skin carotenoid concentrations by the middle of phase 4. Skin carotenoid status predicted plasma values by using a mixed linear model including all time points (r = 0.72, P , 0.001), which indicates that changes in skin carotenoid status closely follow changes in plasma across a broad range of intakes. At the individual level, skin carotenoids predicted plasma values (r = 0.70, P , 0.001) over all time points. Conclusion: Skin carotenoid status assessed by resonance Raman spectroscopy is a noninvasive, objective biomarker of changes in vegetable and fruit intake. This trial was registered at clinicaltrials.gov as NCT01403844.Am J Clin Nutr 2014;100:930-7.