Biomarkers could provide a more accurate measure of long-term intake than questionnaires. Adipose tissue is considered the best indicator of long-term essential fatty acid intake, but other tissues may prove equally valid. The authors evaluated the ability of fasting whole blood, relative to fasting plasma and adipose tissue, to reflect fatty acid intake. Costa Rican men (n=99) and women (n=101) completed a 135-item food frequency questionnaire and provided adipose tissue and blood samples from 1999 to 2001. Fatty acids were identified by using capillary gas chromatography. Correlation coefficients adjusted for age, sex, and body mass index were calculated. Diet-tissue correlation coefficients for α-linolenic acid and linoleic acid, respectively, were 0.38 and 0.43 in whole blood, 0.51 and 0.52 in adipose tissue, and 0.39 and 0.41 in plasma. High correlations were observed between whole-blood α-linolenic and linoleic acid and adipose tissue (r=0.59 and r=0.67) and plasma (r=0.96 and r=0.88), respectively. Results show that fasting whole blood is a suitable biomarker of long-term essential fatty acid intake, and its performance is comparable to that of fasting plasma. Thus, fasting whole blood could be the sample of choice in epidemiologic studies because of its ability to predict intake, its accessibility, and minimum sample processing.