Temperature is one of the most fundamental parameters for the characterization of a physical system. With rapid development of lab-on-a-chip and biology at single cell level, a great demand has risen for the temperature sensors with high spatial, temporal, and thermal resolution. Nevertheless, measuring temperature in liquid environment is always a technical challenge. Various factors may affect the sensing results, such as the fabrication parameters of built-in sensors, thermal property of electrical insulating layer, and stability of fluorescent thermometers in liquid environment. In this review, we focused on different kinds of micro/nano-thermometers applied in the thermal sensing for microfluidic systems and cultured cells. We discussed the advantages and limitations of these thermometers in specific applications and the challenges and possible solutions for more accurate temperature measurements in further studies.