Recently, a new "super" genotype of oats (CDC SO-I or SO-I) has been developed. The objectives of this study were to determine structural makeup (features) of oat grain in endosperm and pericarp regions and to reveal and identify differences in protein amide I and II and carbohydrate structural makeup (conformation) between SO-I and two conventional oats (CDC Dancer and Derby) grown in western Canada in 2006, using advanced synchrotron radiation based Fourier transform infrared microspectroscopy (SRFTIRM). The SRFTIRM experiments were conducted at National Synchrotron Light Sources, Brookhaven National Laboratory (NSLS, BNL, U.S. Department of Energy). From the results, it was observed that comparison between the new genotype oats and conventional oats showed (1) differences in basic chemical and protein subfraction profiles and energy values with the new SO-I oats containing lower lignin (21 g/kg of DM) and higher soluble crude protein (530 g/kg CP), crude fat (59 g/kg of DM), and energy values (TDN, 820 g/kg of DM; NE(L3x), 7.8 MJ/kg of DM); (2) significant differences in rumen biodegradation kinetics of dry matter, starch, and protein with the new SO-I oats containing lower EDDM (638 g/kg of DM) and higher EDCP (103 g/kg of DM); (3) significant differences in nutrient supply with highest truly absorbed rumen undegraded protein (ARUP, 23 g/kg of DM) and total metabolizable protein supply (MP, 81 g/kg of DM) from the new SO-I oats; and (4) significant differences in structural makeup in terms of protein amide I in the endosperm region (with amide I peak height from 0.13 to 0.22 IR absorbance unit) and cellulosic compounds to carbohydrate ratio in the pericarp region (ratio from 0.02 to 0.06). The results suggest that with the SRFTIRM technique, the structural makeup differences between the new genotype oats (SO-I) and two conventional oats (Dancer and Derby) could be revealed.