Mountain lakes are unique and often isolated freshwater habitats that harbour a rich biotic diversity. This high conservation value may be reflected by diatoms, a group of algae that is known for its reliability as a bioindicator, but which has not been studied extensively in mountain lakes of the northern European Alps. In this study, the conservation value of these lakes was assessed by characterizing the number, share, and abundance of diatom Red List (RL) taxa and their relationship with environmental variables, diatom α and β diversity (assemblage uniqueness). For this purpose, linear regression models, generalized linear models, and generalized additive models were fitted and spatial descriptors were included when relevant. Of the 560 diatom taxa identified, 64% were on the RL and half of these were assigned a threat status. As hypothesized, a decreasing share of RL species in sediment and littoral samples at higher trophic levels was reflected by higher total phosphorous content and lower Secchi depth, respectively. Species-rich lakes contained a high number of RL taxa, contrasting our hypothesis of a logarithmic relationship. In turn, RL abundance increased with uniqueness, confirming our initial hypothesis. However, some of the most unique sites were degraded by fish stocking and contained low abundances of RL species. The results demonstrate the importance of oligotrophic mountain lakes as habitats for rare freshwater biota and their vulnerability in light of human impact through cattle herding, tourism, damming, and fish stocking. Additional conservation efforts are urgently needed for mountain lakes that are still underrepresented within legal conservation frameworks. Species richness and uniqueness reflect complementary aspects of RL status and thus should be applied jointly. Uniqueness can indicate both pristine and degraded habitats, so that including information on human impacts facilitates its interpretation.