The present research aims to record the diversity of testate amoebae occurring in different biotopes around Nainital Lake, Uttarakhand, concerning distribution. Free-living, testate amoebae are single-celled protists with external shells and acting excellent bio-indicators of environmental changes. Collection of Tree, Rock and wall Moss samples was done at different locations around the Nainital Lake area from both the littoral zones and the terrestrial regions. The results show an impressive number of species of testate amoebae; 39 species of testate amoebae span over 11 genera and 8 families that all are new for the region and stress the role of some species in biomonitoring and evaluation of the condition of the environment in these peculiar biotopes. This study not only increases researcher’s knowledge on the testate amoebae species distribution in Nainital Lake but also contributes to the body of knowledge that would be useful for the ecological and environmental status of the area in the future. The outcomes of the research confirm the need for the conservation of the habitats in the area surrounding Nainital Lake to sustain the biodiversity and ecosystems of the region.