Chemists and chemistry educators have long sought meaningful ways to visualize fundamentally abstract components, such as atoms and molecules, of their trade. As technology has improved, computer-based visualization methods have infused both research and education in chemistry. Biochemistry, in particular, has become highly dependent on ways that large molecular systems can be represented, and how to best focus attention on the most critical aspects of the molecular system. To better understand the current state of educational efforts related to visual literacy, a needs assessment was developed and administered to a national sample of biochemistry instructors at four-year institutions (N = 536) to determine the types of representations used during biochemistry course instruction and assessment. Cluster analysis was conducted on the responses to determine similar usage of representations in both instruction and assessment. A significant difference was determined between the types of representations used by instructors teaching a biochemistry survey course and a yearlong course. Implications of how these findings can influence biochemistry instruction and assessment are discussed.