The hot springs are known to host a variety of organisms, such as Cyanobacteria, Archaea, and Eukaryotes. The growth and survival of these life forms in extreme environments, where spring water temperature and associated minerals play a significant role, provide analogous conditions like Mars and thus attract researchers to find the possible existence of life beyond the Earth. Many studies have therefore been conducted from the hot springs to understand the controlling factors for these organisms' survival, mainly Cyanobacteria, which are believed to be true thermophiles. However, little is known about diatoms, especially from the hot springs of India, as most of the studies have concentrated on the diversity and distribution of Cyanobacteria. Here, we present a study of diatoms using a geothermal vent sinter from the Puga hot spring of Ladakh, India. Our results suggest that the diatoms preserved in the geothermal vent sinter are less abundant with low diversity and, therefore, represented by a few species only. By correlating the ecological preferences of diatom species with the sinter's morphological characteristics and geochemical analyses, we conclude that these diatom species could be manifested through a secondary deposit on the geothermal vent sinter from the adjacent cold waters of the Puga hot spring. We propose that the temperature gradient could be a key parameter for the occurrence and survival of diatoms in the Puga geothermal vent sinter rather than the gushed hot water. Consequently, the mere presence of diatoms around the hot spring vent cannot be directly linked to their survival in extreme, i.e., hot water conditions. Therefore, eukaryotic forms like diatoms from the hot springs should be used with caution to elucidate the existence of life in extreme (hot water) conditions. In contrast, cold conditions around the hot spring may be the primary drivers for diatoms' survival, which can be used to infer astrobiological implications.