With the U.S. fuel ethanol industry projected to grow during the next several years, supplies of distillers dried grains with solubles (DDGS) are anticipated to continue to grow as well. DDGS is used primarily as livestock feed. Much of the DDGS must be shipped, often over large distances, outside the Corn Belt (which is where most of the corn-based ethanol plants are currently located). Stickiness and caking among particles is a common issue for DDGS, and it often leads to flowability problems. To address this, the objective of this study was to understand the cross-sectional and surface natures of DDGS particles from five ethanol plants, and how they interact with DDGS properties. This study examined the distribution patterns of chemical components within cross-sections, within section edges (i.e., surface layers), and on surfaces using standard staining techniques; chemical composition was determined using standard protocols; and physical and flowability properties were also determined. Crude protein in the samples was 28.33–30.65% db, crude fat was 9.40–10.98% db, and neutral detergent fiber (NDF) was 31.84–39.90% db. Moisture contents were 4.61–8.08% db, and geometric mean diameters were 0.37–0.52 mm. Cross-sectional staining showed protein levels of 19.57–40.39%, and carbohydrate levels of 22.17–43.06%, depending on the particle size examined and the production plant from which the DDGS was sampled. Staining of DDGS particles indicated a higher amount of surface layer protein compared with carbohydrate thickness in DDGS particles that had a lower flow function index (which indicated potential flow issues). Additionally, surface fat staining suggested that higher surface fat also occurred in samples with worse flow problems. This study represents another step toward understanding why DDGS particles stick together during storage and transport, and will hopefully help to improve DDGS material handling strategies.