Vegetation, through its condition, reflects the properties of the environment. Heterogeneous alpine ecosystems play a critical role in global monitoring systems, but due to low accessibility, cloudy conditions, and short vegetation periods, standard monitoring methods cannot be applied comprehensively. Hyperspectral tools offer a variety of methods based on narrow-band data, but before extrapolation to an airborne or satellite scale, they must be verified using plant biometrical variables. This study aims to assess the condition of alpine sward dominant species (Agrostis rupestris, Festuca picta, and Luzula alpino-pilosa) of the UNESCO Man&Biosphere Tatra National Park (TPN) where the high mountain grasslands are strongly influenced by tourists. Data were analyzed for trampled, reference, and recultivated polygons. The field-obtained hyperspectral properties were verified using ground measured photosynthetically active radiation, chlorophyll content, fluorescence, and evapotranspiration. Statistically significant changes in terms of cellular structures, chlorophyll, and water content in the canopy were detected. Lower values for the remote sensing indices were observed for trampled plants (about 10â15%). Species in recultivated areas were characterized by a similar, or sometimes improved, spectral properties than the reference polygons; confirmed by fluorescence measurements (Fv/Fm). Overall, the fluorescence analysis and remote sensing tools confirmed the suitability of such methods for monitoring species in remote mountain areas, and the general condition of these grasslands was determined as good.