In vivo O₂ imaging in hepatic tissues by phosphorescence lifetime imaging microscopy using Ir(III) complexes as intracellular probes.

ABSTRACT: Phosphorescence lifetime imaging microscopy (PLIM) combined with an oxygen (O₂)-sensitive luminescent probe allows for high-resolution O₂ imaging of living tissues. Herein, we present phosphorescent Ir(III) complexes, (btp)₂Ir(acac-DM) (Ir-1) and (btp-OH)₃Ir (Ir-2), as useful O₂ probes for PLIM measurement. These small-molecule probes were efficiently taken up into cultured cells and accumulated in specific organelles. Their excellent cell-permeable properties allowed for efficient staining of three-dimensional cell spheroids, and thereby phosphorescence lifetime measurements enabled the evaluation of the O₂ level and distribution in spheroids, including the detection of alterations in O₂ levels by metabolic stimulation with an effector. We took PLIM images of hepatic tissues of living mice by intravenously administrating these probes. The PLIM images clearly visualized the O₂ gradient in hepatic lobules with cellular-level resolution, and the O₂ levels were derived based on calibration using cultured cells; the phosphorescence lifetime of Ir-1 gave reasonable O₂ levels, whereas Ir-2 exhibited much lower O₂ levels. Intravenous administration of NH₄Cl to mice caused the hepatic tissues to experience hypoxia, presumably due to O₂ consumption to produce ATP required for ammonia detoxification, suggesting that the metabolism of the probe molecule might affect liver O₂ levels.

SUBMITTER: Mizukami K 

PROVIDER: S-EPMC7713648 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature