Poor nutrition and landscape changes are regularly cited as key factors causing the decline of wild and managed bee populations. However, what constitutes “poor nutrition” for bees currently is inadequately defined. Bees collect and eat pollen: it is their only solid food source and it provides a broad suite of required macro- and micronutrients. Bees are also generalist foragers and thus the different pollen types they collect and eat can be highly nutritionally variable. Therefore, characterizing the multidimensional nutrient content of different pollen types is needed to fully understand pollen as a nutritional resource. Unfortunately, the use of different analytical approaches to assess pollen nutrient content has complicated between-studies comparisons and blurred our understanding of pollen nutrient content. This dataset includes the raw data generated from each of our analyses, including pollen disruption and nutrient assays. We used our collective data to propose a unifying approach for analyzing pollen nutrient content. This will help researchers better study and understand the nutritional ecology – including foraging behavior, nutrient regulation, and health – of bees and other pollen feeders.