In vivo O(2) imaging in hepatic tissues by phosphorescence lifetime imaging microscopy using Ir(III) complexes as intracellular probes.
In vivo O(2) imaging in hepatic tissues by phosphorescence lifetime imaging microscopy using Ir(III) complexes as intracellular probes.
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DOI：
10.1038/s41598-020-76878-6
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发表时间：
2020-12-03
影响因子：
4.6
通讯作者：
Tobita S
中科院分区：
文献类型：
--
作者：
Mizukami K;Katano A;Shiozaki S;Yoshihara T;Goda N;Tobita S
磷光寿命成像显微镜（PLIM）与氧（O2）敏感的发光探针相结合，允许活组织的高分辨率O2成像。本文中，我们提出磷光Ir（III）配合物，（btp）2 Ir（acac-DM）（Ir-1）和（btp-OH）3 Ir（Ir-2），作为PLIM测量的有用的O2探针。这些小分子探针被有效地吸收到培养的细胞中，并在特定的细胞器中积累。其优异的细胞可渗透性允许三维细胞球体的有效染色，从而磷光寿命测量能够评价球体中的O2水平和分布，包括通过效应物的代谢刺激检测O2水平的改变。我们通过静脉注射这些探针对活体小鼠的肝组织进行PLIM成像。PLIM图像清楚地显示了具有细胞水平分辨率的肝小叶中的O2梯度，并且O2水平是基于使用培养细胞的校准导出的; Ir-1的磷光寿命给出了合理的O2水平，而Ir-2表现出低得多的O2水平。小鼠静脉注射NH 4Cl导致肝组织缺氧，可能是由于O2消耗产生氨解毒所需的ATP，这表明探针分子的代谢可能会影响肝脏O2水平。
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.
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影响因子：
2
作者：
Felder AE;Wanek J;Teng PY;Blair NP;Shahidi M
通讯作者：
Shahidi M
影响因子：
3.6
作者：
Liu, Y. L.;Yan, T.;Yang, X. J.
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Yang, X. J.
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Arden, Geoffrey B.;Sivaprasad, Sobha
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Sivaprasad, Sobha
影响因子：
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Nichols, Alexander J.;Roussakis, Emmanuel;Klein, Oliver J.;Evans, Conor L.
通讯作者：
Evans, Conor L.
影响因子：
6.6
作者：
Dmitriev, Ruslan I.;Kondrashina, Alina V.;Papkovsky, Dmitri B.
通讯作者：
Papkovsky, Dmitri B.