Liver and skeletal muscle triglyceride stores are elevated in type 2 diabetes and correlate with insulin resistance. As postprandial handling of dietary fat may be a critical determinant of tissue triglyceride levels, we quantified postprandial fat storage in normal and type 2 diabetes subjects. Healthy volunteers (n = 8) and diet-controlled type 2 diabetes subjects (n = 12) were studied using a novel ¹³C magnetic resonance spectroscopy protocol to measure the postprandial increment in liver and skeletal muscle triglyceride following ingestion of ¹³C-labeled fatty acids given with a standard mixed meal. The postprandial increment in hepatic triglyceride was rapid in both groups (peak increment controls: +7.3 ± 1.5 mmol/l at 6 h, P = 0.002; peak increment diabetics: +10.8 ± 3.4 mmol/l at 4 h, P = 0.009). The mean postprandial incremental AUC of hepatic ¹³C enrichment between the first and second meals (0 and 4 h) was significantly higher in the diabetes group (6.1 ± 1.4 vs. 1.7 ± 0.6 mmol·l⁻¹·h⁻¹, P = 0.019). Postprandial increment in skeletal muscle triglyceride in the control group was small compared with the diabetic group, the mean 24-h postprandial incremental AUC being 0.2 ± 0.3 vs. 1.7 ± 0.4 mmol·l⁻¹·h⁻¹ (P = 0.009). We conclude that the postprandial uptake of fatty acids by liver and skeletal muscle is increased in type 2 diabetes and may underlie the elevated tissue triglyceride stores and consequent insulin resistance.