Bias in Intracellular Luminescence Thermometry: The Case of the Green Fluorescent Protein

Measurement of intracellular temperature in a fast, accurate, reliable, and remote manner is crucial for the understanding cellular processes. Nanothermometers based on green fluorescence protein (GFP) are special interest because readouts can be obtained from analysis polarization state its luminescence. Despite good results provided by GFP thermometers, reliability their thermal still question debate. Here, light shed this issue introducing cell activity as relevant bias mechanism that prevents use reliable intranuclear measurements. Experimental evidence lack affect not only but also other widely used thermometers such semiconductor nanocrystals provided. It discussed how differences observed between calibration curves presence absence inform about bias. The presented discussion aimed to warn community working thermometry encourage authors approach conscious manner. performance chosen must judiciously assessed. This way progress deliver indisputable results.