Abstract Carotenoid pigments play an important role in the seasonal regulation of photosynthesis and photoprotection of overwintering conifers. Because the seasonal changes in the rate of photosynthetic CO2 assimilation are linked to changes in carotenoid pigment composition, it has been suggested that carotenoid sensitive vegetation indices might be used to track the otherwise “invisible” phenology of photosynthesis of conifer forests through remote sensing of leaf spectral reflectance. In this study we aimed to assess differences in the seasonal regulation of photosynthesis and the associated variation of carotenoids and chlorophylls at the leaf-scale for eastern white pine, red maple and white oak, in order to understand if photosynthetic and photoprotective processes are adequately represented by different vegetation indices over the course of the year. For this purpose we measured maximum rates of CO2 assimilation (Amax), quantified photosynthetic pigments, estimated photochemical and non-photochemical quenching processes via chlorophyll fluorescence and determined leaf spectral reflectance in pine, maple and oak trees over the course of two years. Seasonal variation in Amax, used here as a proxy for photosynthetic phenology, and photosynthetic pigments were adequately represented by the normalized difference vegetation index (NDVI) for the deciduous trees. For pine, NDVI overestimated photosynthetic activity for most of the year and was hence not able to represent photosynthetic phenology, due to the fact that needle chlorophyll content shows only little variation over the course of the year. By contrast, using the photochemical reflectance index (PRI) and the chlorophyll/carotenoid index (CCI), which both detect variations in carotenoids, we were able to observe an improved representation of the seasonal variation of CO2 assimilation and photosynthetic phenology for the two deciduous and the conifer species. Based on the accurate detection of the seasonal regulation of leaf-scale photosynthetic activity for all three species, we conclude that carotenoid-sensitive vegetation indices are promising tools to improve monitoring of phenology in both deciduous and conifer forests.