Ir(iii) complex-based oxygen imaging of living cells and ocular fundus with a gated ICCD camera
Ir(iii) complex-based oxygen imaging of living cells and ocular fundus with a gated ICCD camera
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DOI：
10.1039/c8pp00122g
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发表时间：
2018-06-01
影响因子：
3.1
通讯作者：
Tobita, S.
中科院分区：
文献类型：
--
作者：
Akiyama, H.;Takahashi, I.;Tobita, S.
使用氧敏感探针的磷光寿命成像方法对于高空间分辨率地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磷光氧探针和最佳检测方法的有用的氧检测技术。本文提出了一种生物氧成像方法，使用配有门控强化电荷耦合器件（ICCD）相机的显微镜作为探测器，Ir（iii）配合物作为磷光氧探针。使用细胞穿透Ir（iii）复合物BTPDM1和倒置显微镜获得的单层HT-29细胞（人结直肠腺癌细胞）的显微发光图像表明，当单层被薄盖盖住时，这种方法可以可视化培养细胞单层中产生的氧梯度。此外，将红外发射Ir（iii）复合物DTTPH-PEG（24）与配备门控ICCD相机的微缩显微镜结合，既可以显示视盘附近的视网膜血管，又可以监测兔视网膜吸入氧含量变化时的氧水平变化。
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-PEG(24) 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.