Measurement of Intracellular Temperature in Brown Adipocytes Using a Cationic Fluorescent Polymeric Thermometer

Brown adipose tissue specializes in expending energy through non-shivering thermogenesis, and many studies have associated its activity with protection and treatment of obesity and metabolic diseases. To reveal the mechanisms involved in heat production, primary cultured brown adipose cells (BACs) have been used because of their ease of genetic engineering and similarity to living tissue. However, thermogenic activity has often been evaluated as an indirect method, such as the measurement of oxygen consumption. Recently, fluorescent nanothermometers for the direct measurement of intracellular temperature have been developed and applied to elucidate the mechanisms of heat production in BACs. In this chapter, we introduce a protocol that uses a cationic fluorescent polymeric thermometer to directly measure the temperature within primary cultured BACs. We anticipate that this protocol will be beneficial in elucidating the mechanism of thermogenesis in BACs.