The outburst of superoxide anion (O₂⁻) in mitochondrial during ischemia/reperfusion (I/R) process will cause a series of oxidative damage including polarity loss of mitochondrial membrane potential, overload of secondary cellular calcium, and cascade apoptosis. To monitor the O₂⁻ level fluctuations as well as to evaluate the relationship between O₂⁻ concentration and the degree of cell apoptosis during I/R process, we propose a ratiometric near-infrared mitochondrial targeting fluorescent probe Mito-Cy-Tfs for the detection of level changes of O₂⁻ in cells and in vivo. The probe Mito-Cy-Tfs is composed of three moieties: near-infrared heptamethine cyanine as fluorescence signal transducer, trifluoromethanesulfonamide as fluorescence modulator, and lipophilic triphenylphosphonium cation as mitochondrial guider. The probe can well locate in mitochondria and respond the concentration changes of endogenous O₂⁻ selectively and sensitively. The probe has been successfully utilized to image the endogenous O₂⁻ fluctuations in four kinds of cell I/R models (glucose deprivation/reperfusion, serum deprivation/reperfusion, oxygen deprivation/reperfusion and glucose-serum-oxygen deprivation/reperfusion). The probe also exhibits deep tissue penetration for real-time imaging of O₂⁻concentration in liver of I/R mice model. We confirm that the adoption of ischemic preconditioning (IPC) and postconditioning (IPTC) can protect liver from I/R injury. The probe can be employed to accurately indicate and evaluate the mutual relationship between the levels of O₂⁻ and the degrees of organ damage during I/R, IPC and IPTC processes. The above applications make our new probe a potential candidate for the clinical surgery assessment.

超氧阴离子（O ₂⁻）在线粒体缺血/再灌注（I / R）过程中会导致一系列氧化损伤，包括线粒体膜电位的极性损失，继发性细胞钙超载和级联细胞凋亡。监视O ₂⁻液位波动以及评估O ₂之间的关系⁻在I / R过程中浓度和细胞凋亡程度，我们提出了一种比例式近红外线粒体靶向荧光探针Mito-Cy-Tfs，用于检测O ₂的水平变化⁻在细胞和体内。Mito-Cy-Tfs探针由三个部分组成：近红外七甲基花青作为荧光信号转导子，三氟甲磺酰胺作为荧光调节剂，亲脂性三苯基phosph阳离子作为线粒体引导子。该探针可以很好地定位于线粒体中并响应内源性O ₂的浓度变化⁻有选择地和敏感地。该探针已成功地用于对内源性O ₂进行成像⁻四种细胞I / R模型的波动（葡萄糖剥夺/再灌注，血清剥夺/再灌注，氧气剥夺/再灌注和葡萄糖-血清氧气剥夺/再灌注）。该探头还具有深层组织穿透性，可对O ₂进行实时成像⁻ I / R小鼠模型在肝脏中的浓度。我们确认采用缺血预处理（IPC）和后处理（IPTC）可以保护肝脏免受I / R损伤。该探针可用于准确指示和评估O ₂水平之间的相互关系⁻以及在I / R，IPC和IPTC过程中器官受损的程度。上述应用使我们的新探针成为临床手术评估的潜在候选者。