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应用低能扫描透射电子显微镜研究 Pt 纳米颗粒在人结肠癌细胞中的摄取。

Application of low-energy scanning transmission electron microscopy for the study of Pt-nanoparticle uptake in human colon carcinoma cells.

机构信息

Laboratorium für Elektronenmikroskopie, Karlsruher Institut für Technologie (KIT) , 76128 Karlsruhe , Germany.

出版信息

Nanotoxicology. 2014 Jun;8(4):433-46. doi: 10.3109/17435390.2013.796535. Epub 2013 May 9.

Abstract

High-angle annular dark-field scanning transmission electron microscopy (HAADF STEM) in a scanning electron microscope facilitates the acquisition of images with high chemical sensitivity and high resolution. HAADF STEM at low electron energies is particularly suited to image nanoparticles (NPs) in thin cell sections which are not subjected to poststaining procedures as demonstrated by comparison with bright-field TEM. High membrane contrast is achieved and distinction of NPs with different chemical composition is possible at first sight. Low-energy HAADF STEM was applied to systematically study the uptake of Pt-NPs with a broad size distribution in HT29 colon carcinoma cells as a function of incubation time and incubation temperature. The cellular dose was quantified, that is, the amount and number density of NPs taken up by the cells, as well as the particle-size distribution. The results show a strong dependence of the amount of incubated NPs on the exposure time which can be understood by considering size-dependent diffusion and gravitational settling of the NPs in the cell culture medium.

摘要

高角环形暗场扫描透射电子显微镜(HAADF-STEM)在扫描电子显微镜中,有利于获取具有高化学灵敏度和高分辨率的图像。在低电子能量下的 HAADF-STEM 特别适合于对薄细胞切片中的纳米颗粒(NPs)进行成像,这些切片不需要进行后染色处理,这一点通过与明场 TEM 的比较得到了证明。可以实现高膜对比度,并可以一眼区分具有不同化学成分的 NPs。低能 HAADF-STEM 被应用于系统地研究具有较宽尺寸分布的 Pt-NPs 在 HT29 结肠癌细胞中的摄取,作为孵育时间和孵育温度的函数。细胞剂量被量化,即被细胞摄取的 NPs 的数量和数密度,以及颗粒尺寸分布。结果表明,孵育的 NPs 的数量强烈依赖于暴露时间,这可以通过考虑 NPs 在细胞培养液中的尺寸依赖性扩散和重力沉降来理解。

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