In this work, the response of temperate coniferous forests to ozone air pollution (O3) in the mountain environment of the High Tatra Mts. (Western Carpathians) was analyzed. The modelling of stomatal O3 flux is a complex method for the estimation of phytotoxicity of O3 pollution to forest vegetation. Stomatal flux-based critical levels (CLef) for effects of O3 on radial growth take into account the varying influences of O3 concentration, meteorological variables, soil properties, and phenology. The application of the model DO3SE (Deposition of Ozone for Stomatal Exchange) at five experimental plots with altitudes varying from 810 to 1,778 m a.s.l. along vertical and spatial profile in the High Tatra Mts. revealed the high phytotoxic potential of O3 on spruce forests during the growing season 2014. The accumulated stomatal O3 flux above a threshold of Y (1 nmol m−2 s−1), i.e. POD1 (Phytotoxic Ozone Dose) ranged from 13.6 mmol m−2 at the Kolové pleso site (1,570 m a.s.l.) to 16.2 mmol m−2 at Skalnaté Pleso site (1,778 m a.s.l.). CLef for POD1 (8 mmol m−2) recommended for the protection of spruce forests were exceeded at all experimental plots from early July. Similarly, AOT40 index suggests vulnerability of mountain forests to O3 pollution. AOT40 values increased with altitude and reached values varying from 6.2 ppm h in Stará Lesná (810 m a.s.l.) to 10.7 ppm h at Skalnaté Pleso close to the timber line (1,778 m a.s.l.). Concentration-based critical level (CLec) of 5,000 ppb h was exceeded from June to August and was different for each experimental site.