Improved nitrogen use efficiency (NUE) in crop plants has the potential to significantly reduce fertilizer application costs and increase crop yield. NUE is a complex, multi-component trait and screening for NUE in field trials poses the challenge of low heritability and high environmental interaction. High resolution image based phenotyping technologies in controlled environments allow for the non-destructive measurement of plant growth and is a useful tool for determining nitrogen (N) response. However, growth response by itself does not tell us anything about the dynamics of uptake, allocation and remobilisation. Here we present results from using a high throughput phenotyping platform to investigate the nitrogen response of two wheat varieties combined with hyperspectral based non-destructive measurement of leaf N content to determine N dynamics. Treatments incorporated N additions during growth to reflect common farmer practice. In terms of biomass there were clear differences in N response between high and low N treatments, including those with supplemented N. Tissue N showed a general pattern of increase then decrease during the life of a leaf but overall levels differed between treatments and varieties. The N addition treatments showed dramatic increases in leaf N in a short time frame. Being able to measure the dynamics of plant responses to N over time, combined with imaged based growth analysis may greatly enhance our chances to develop high NUE cereals.