To satisfy emerging energy demands, cellulosic biomass shows the potential to improve U.S. energy security and lower greenhouse gases emissions. Cellulosic biomass is valuable and has manifold possible uses, such as manufacturing advanced biofuels, supplying materials for biomass burners and power plants. Growing cellulosic biomass on non-agricultural lands, such as highway right-of-way (ROW), can avoid land use competition between energy and food crops on agricultural lands. Several Departments of Transportation around the U.S. have considered this opportunity because of typically millions of dollars expenditures for mowing and maintaining costs every year. Growing energy crops, while replacing turf grasses, provides opportunity to create economic benefits. This study will analyze the optimal harvesting and logistics operation strategies for highway ROW scenarios for Illinois Department of Transportation (IDOT). Available lands for growing energy crops along highway ROW can be identified and grouped into segments by Geographic Information System (GIS) data analysis. Given the information regarding available lands, biomass feedstock production optimization modeling is applied to determine harvest schedules and estimate biomass yield. Expected results will identify the optimal trip assignment and truck routing strategies for the IDOT scenarios. Estimated operational cost and time are also calculated. These results will provide information for decision makers and equipment operators, not only determining the best operational strategy, but also keeping flexibility for the realistic application.