Phosphorescence lifetime imaging methods using oxygen-sensitive probes are very useful for visualizing the oxygen status of living cells and tissues with high spatial resolution. We aim to develop a useful oxygen detection technique combining a phosphorescent oxygen probe and an optimal detection method. Herein we present a biological oxygen imaging method using a microscope equipped with a gated intensified charge-coupled device (ICCD) camera as a detector and an Ir(III) complex as a phosphorescent oxygen probe. Microscopic luminescence images of monolayer HT-29 cells (human colorectal adenocarcinoma cells) obtained using the cell-penetrating Ir(III) complex BTPDM1 and an inverted microscope demonstrated that this method allowed visualization of the oxygen gradient produced in a monolayer of cultured cells when the monolayer is covered with a thin coverslip. Furthermore, combining the IR-emitting Ir(III) complex DTTPH-PEG₂₄ with a macrozoom microscope equipped with a gated ICCD camera enabled both the visualization of retinal vessels near the optic disc and the monitoring of oxygen level changes in a rabbit retina upon changing the inhaled oxygen content.

中文翻译：

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带门ICCD摄像机的活细胞和眼底基于 Ir（iii）配合物的氧成像†

使用氧敏感探针的磷光寿命成像方法对于以高空间分辨率可视化活细胞和组织的氧状态非常有用。我们的目标是开发一种结合了磷光氧气探针和最佳检测方法的有用的氧气检测技术。本文中，我们介绍了一种生物氧气成像方法，该方法使用配备有门控增强型电荷耦合器件（ICCD）相机作为检测器和Ir（III）配合物作为磷光氧探针的显微镜。使用穿透细胞的Ir（III）获得的单层HT-29细胞（人结肠直肠腺癌细胞）的显微发光图像）复杂的BTPDM1和倒置显微镜证明，当单层薄薄的盖玻片覆盖单层培养细胞时，该方法可以观察到培养细胞单层中产生的氧梯度。此外，将发射红外光的Ir（III）复合物DTTPH-PEG ₂₄与配备门控ICCD相机的大型变焦显微镜相结合，不仅可以使视盘附近的视网膜血管可视化，还可以监测兔子视网膜改变后的氧气水平变化吸入的氧气含量。