The strong correlation between chlorophyll fluorescence (ChlF) parameters and photosynthesis highlights the need for a comprehensive spatial representation of ChlF parameters within the canopy. Such an approach is essential to advance our understanding and to improve the representation and modeling of water and carbon fluxes at scales ranging from the leaf to the canopy level. However, the challenge remains in determining how to effectively describe and track the variability of ChlF parameters within the canopy. In this study, we determined the variation in leaf biophysical and chemical traits and ChlF parameters along the vertical height of the canopy for several species in a temperate deciduous forest. We observed general associations of height with leaf biophysical and chemical traits and ChlF parameters, although these relationships were species‐dependent. In addition, leaf biophysical and chemical traits, particularly light‐harvesting pigments, showed significant effects on ChlF parameters. To effectively track variation in ChlF parameters within the canopy, we used gradient‐boosted regression (GBR) models driven by leaf traits and species, which explained more than 80% of the variation in all ChlF parameters. Our study demonstrates the feasibility of utilizing leaf biophysical and chemical traits to predict vertical variation in ChlF parameters and provide supportive data for modeling canopy photosynthesis.