Decline of physical function [PF] in old age might be related to oxidative damage caused by free radicals, and antioxidants may play a role in reducing the risk of physical functional limitations [PFL]. Yet little is known about the role of carotenoids in PFL. The purpose of this study was to assess the association of total and daily dietary intakes of carotenoids, fruit and vegetables [FV] and their biomarkers with PF among U.S. older adults. Data were from 2,905 men and women [≥ 60 years] in the National Health and Nutrition Examination Survey [NHANES] 2003-2006. Using logistic regression, we found that serum concentration of carotenoids was associated with limitations in PF. In the fully adjusted model, the ORs [95% CI] of having limitation in activities of daily living [ADLs], instrumental activities of daily living [IADLs] and movement difficulties [MD] were 2.03 [1.16 - 3.53], 2.34 [1.61 - 3.42], and 2.15 [1.46 - 3.18], respectively, comparing the lowest quintile of serum carotenoids to the highest. Total intake and dietary intake of carotenoids were found to be associated with limitations in IADL. However, low FV consumptions were not significantly associated with PF domains. In conclusion, elevated levels of serum carotenoids are significantly associated with better physical functional performance and may play an important role in delaying the onset of physical decline.