近红外荧光探针用于细胞内单线态氧的检测及成像分析
首发时间:2012-11-30
摘要:单线态氧(1O2)是重要的活性氧物种,并且是一种强的氧化剂。它能与许多生物分子如脂类、核酸、氨基酸、多肽和蛋白质等反应,引起机体组织损伤。另一方面,1O2又可以控制、调节机体功能,是生物体免疫系统的主要杀菌剂,并已用于癌症的光动力治疗。因此,生物体内1O2的有效检测对研究1O2的分子作用机制十分重要。我们以近红外荧光染料花菁为母体,以蒽基为特异性识别基团合成了一种检测1O2的近红外荧光探针An-Cy。探针本身荧光很弱,跟1O2反应后荧光显著增强,探针的激发和发射波长分别为740 nm和805 nm,检测1O2的线性范围为1.0×10-7~ 3.0×10-6 M,线性方程为ΔF= 877.7+ 443.0[1O2]( μM),线性相关系数为0.9928,检测限为51 nM。运用激光共聚焦荧光成像技术,成功实现了小鼠巨噬细胞(RAW 264.7)线粒体中1O2的荧光定位成像。研究结果为细胞及生物体中1O2的定位分析提供了理想的荧光探针和分析方法。
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Design and synthesis of a near-infrared fluorescent probe for singlet oxygen in living cells
Abstract:The design and synthesis of a near-infrared fluorescent probe (An-Cy) for the detection and imaging of singlet oxygen in living cells is accomplished. A near-infrared fluorescent probe An-Cy that synthesized through combining anthryl group and cyanine is proved to be a turn-on type probe for singlet oxygen. The initial fluorescence of An-Cy is low, but is significantly enhanced in the presence of singlet oxygen (λex/λem=740/805 nm). The standard equation is ΔF= 877.7+ 443.0[1O2] (μM) with the linear range and detection limit of 1.0×10-7~3.0×10-6 M and 51 nM, respectively. Using confocal fluorescence imaging, the singlet oxygen that produced in the PMA-stimulated murine macrophages (RAW 264.7) is analyzed successfully. The results provide an ideal fluorescent probe and corresponding analytical method for the analysis of singlet oxygen in living cells and organisms, and exhibits important practical significance.
Keywords: analytical chemistry singlet oxygen near-infrared fluorescent probe confocal fluorescence imaging
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