活性氧
纳米毒理学
荧光寿命成像显微镜
氧化应激
生物物理学
活体细胞成像
纳米颗粒
荧光显微镜
纳米技术
荧光
DNA损伤
自体荧光
材料科学
化学
细胞生物学
细胞
生物化学
生物
DNA
物理
量子力学
作者
Jens Balke,Pierre Volz,Falko Neumann,Robert Brodwolf,Alexander L. Wolf,Hannah Pischon,Moritz Radbruch,Lars Mundhenk,Achim D. Gruber,Nan Ma,Ulrike Alexiev
出处
期刊:Small
[Wiley]
日期:2018-06-01
卷期号:14 (23): 1800310-1800310
被引量:20
标识
DOI:10.1002/smll.201800310
摘要
Nanoparticles hold a great promise in biomedical science. However, due to their unique physical and chemical properties they can lead to overproduction of intracellular reactive oxygen species (ROS). As an important mechanism of nanotoxicity, there is a great need for sensitive and high-throughput adaptable single-cell ROS detection methods. Here, fluorescence lifetime imaging microscopy (FLIM) is employed for single-cell ROS detection (FLIM-ROX) providing increased sensitivity and enabling high-throughput analysis in fixed and live cells. FLIM-ROX owes its sensitivity to the discrimination of autofluorescence from the unique fluorescence lifetime of the ROS reporter dye. The effect of subcytotoxic amounts of cationic gold nanoparticles in J774A.1 cells and primary human macrophages on ROS generation is investigated. FLIM-ROX measures very low ROS levels upon gold nanoparticle exposure, which is undetectable by the conventional method. It is demonstrated that cellular morphology changes, elevated senescence, and DNA damage link the resulting low-level oxidative stress to cellular adverse effects and thus nanotoxicity. Multiphoton FLIM-ROX enables the quantification of spatial ROS distribution in vivo, which is shown for skin tissue as a target for nanoparticle exposure. Thus, this innovative method allows identifying of low-level ROS in vitro and in vivo and, subsequently, promotes understanding of ROS-associated nanotoxicity.
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