Dual-Emissive Phosphorescent Polymer Probe for Accurate Temperature Sensing in Living Cells and Zebrafish Using Ratiometric and Phosphorescence Lifetime Imaging Microscopy

Temperature plays an important part in many biochemical processes. Accurate diagnosis and proper treatment usually depend on precise measurement of temperature. In this work, a dual-emissive phosphorescent polymer temperature probe, composed of iridium(III) complexes as temperature sensitive unit with phosphorescence lifetime of ∼500 ns and europium(III) complexes as reference unit with lifetime of ∼400 μs, has been rationally designed and synthesized. Upon the increase of the temperature, the luminescence intensity from the iridium(III) complexes is enhanced, while that from the europium(III) complexes remains unchanged, which makes it possible for the ratiometric detection of temperature. Furthermore, the polymer also displays a significant change in emission lifetime accompanied by the temperature variation. By utilizing the laser scanning confocal microscope and time-resolved luminescence imaging systems, ratiometric and time-resolved luminescence imaging in Hela cells and zebrafish have been carried out. Notably, the intensity ratio and long-lifetime-based imaging can offer higher sensitivity, decrease the detection limit, and minimize the background interference from biosamples.