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利用共聚焦显微镜对活细胞和组织内无标记纳米颗粒进行长期细胞内追踪。

Long-Term Intracellular Tracking of Label-Free Nanoparticles in Live Cells and Tissues with Confocal Microscopy.

机构信息

Institut Català de Nanociència i Nanotecnologia (ICN2), Campus UAB, Bellaterra, Barcelona, 08193, Spain.

Vall d'Hebron Institut of Research (VHIR), Barcelona, 08035, Spain.

出版信息

Small Methods. 2024 Oct;8(10):e2301713. doi: 10.1002/smtd.202301713. Epub 2024 Apr 2.

Abstract

The label-free imaging of inorganic nanoparticles (NPs) using confocal laser scanning microscopy (CLSM) provides a powerful and versatile tool for studying interactions between NPs and biological systems. Without the need for exogenous labels or markers, it simply benefits from the differential scattering of visible photons between biomaterials and inorganic NPs. Validation experiments conducted on fixed and living cells in real-time, as well as mouse tissue sections following parenteral administration of NPs. Additionally, by incorporating reporter fluorophores and utilizing both reflectance and fluorescence imaging modalities, the method enables high-resolution multiplex imaging of cellular structures and NPs. Different sizes and concentrations of Au NPs are tested as for Ag, FeO, and CeO NPs, all with biological interest. Overall, the comprehensive study of NP imaging by confocal microscopy in reflectance mode provides valuable insights and tools for researchers interested in monitoring the nano-bio interactions.

摘要

利用共聚焦激光扫描显微镜(CLSM)对无标记的无机纳米粒子(NPs)进行成像,为研究 NPs 与生物系统之间的相互作用提供了一种强大且多功能的工具。该方法无需外源性标记或标记物,仅利用生物材料与无机 NPs 之间可见光光子的差异散射。在实时固定和活细胞以及 NPs 静脉给药后的小鼠组织切片上进行了验证实验。此外,通过掺入报告荧光团并利用反射和荧光成像模式,该方法可以实现细胞结构和 NPs 的高分辨率多重成像。对具有生物学意义的不同尺寸和浓度的 Au NPs 以及 Ag、FeO 和 CeO NPs 进行了测试。总的来说,通过共聚焦显微镜反射模式对 NP 成像进行的综合研究为关注监测纳米-生物相互作用的研究人员提供了有价值的见解和工具。

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