It has been known that cells have mechanisms to sense and respond to environmental noxiousness and mild temperature changes, such as heat shock response and thermosensitive TRP channels. Meanwhile, new methods of measuring temperature at the cellular level has recently been developed using fluorescent nanothermometers. Among these thermometers, fluorescent polymeric thermometers and fluorescent nanodiamonds excel in the properties required for intracellular thermometry. By using these novel methods to measure the temperature of single cells in cultures and tissues, it was revealed that spontaneous spatiotemporal temperature fluctuations occur within cells. Furthermore, the temperature fluctuations were related to organelles such as mitochondria and cellular and physiological functions, revealing a close relationship between intracellular temperature and cellular functions.