Quantification of silver nanoparticles taken up by single cells using inductively coupled plasma mass spectrometry in the single cell measurement mode†,Journal of Analytical Atomic Spectrometry

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Quantification of silver nanoparticles taken up by single cells using inductively coupled plasma mass spectrometry in the single cell measurement mode†
Journal of Analytical Atomic Spectrometry ( IF 3.1 ) Pub Date : 2018-05-18 00:00:00 , DOI: 10.1039/c7ja00395a
Ana López-Serrano Oliver _{1,

2,

3} , Sabine Baumgart _{2,

3,

4} , Wolfram Bremser _{1,

2,

3} , Sabine Flemig _{1,

2,

3} , Doreen Wittke _{3,

5,

6,

7} , Andreas Grützkau _{2,

3,

4} , Andreas Luch _{3,

5,

6,

7} , Andrea Haase _{3,

5,

6,

7} , Norbert Jakubowski _{1,

2,

3}

Affiliation

The impact of nanoparticles, NPs, at the single cell level has become a major field of toxicological research and different analytical methodologies are being investigated to obtain biological and toxicological information to better understand the mechanisms of cell–NP interactions. Here, inductively coupled plasma mass spectrometry in the single cell measurement mode (SC-ICP-MS) is proposed to study the uptake of silver NPs, AgNPs, with a diameter of 50 nm by human THP-1 monocytes in a proof-of-principle experiment. The main operating parameters of SC-ICP-MS have been optimized and applied for subsequent quantitative analysis of AgNPs to determine the number of particles in individual cells using AgNP suspensions for calibration. THP-1 cells were incubated with AgNP suspensions with concentrations of 0.1 and 1 μg mL⁻¹ for 4 and 24 hours. The results reveal that the AgNP uptake by THP-1 monocytes is minimal at the lower dose of 0.1 μg mL⁻¹ (roughly 1 AgNP per cell was determined), whereas a large cell-to-cell variance dependent on the exposure time is observed for a 10 times higher concentration (roughly 7 AgNPs per cell). The method was further applied to monitor the AgNP uptake by THP-1 cells differentiated macrophages incubated at the same AgNP concentration levels and exposure times demonstrating a much higher AgNP uptake (roughly from 9 to 45 AgNPs per cell) that was dependent on exposure concentration and remained constant over time. The results have been compared and validated by sample digestion followed by ICP-MS analysis as well as with other alternative promising techniques providing single cell analysis.

中文翻译：

在单细胞测量模式下使用电感耦合等离子体质谱法定量单细胞吸收的银纳米颗粒†

纳米颗粒，NPs在单细胞水平上的影响已成为毒理学研究的主要领域，并且正在研究不同的分析方法以获得生物学和毒理学信息，以更好地了解细胞-NP相互作用的机制。在这里，建议采用单细胞测量模式的电感耦合等离子体质谱（SC-ICP-MS），以研究人类THP-1单核细胞对直径为50 nm的银NPs，AgNPs的摄取。原理实验。SC-ICP-MS的主要操作参数已经过优化，可用于随后的AgNP定量分析，以确定使用AgNP悬浮液进行标定的单个细胞中的颗粒数量。将THP-1细胞与浓度为0.1和1μgmL ^-1的AgNP悬浮液孵育持续4到24小时。结果表明，在0.1μgmL ^-1的较低剂量下，THP-1单核细胞摄取的AgNP最小（确定每个细胞大约1 AgNP），而观察到的较大的细胞间差异取决于暴露时间浓度提高了10倍（每个细胞大约7个AgNP）。该方法进一步应用于监测THP-1细胞分化的巨噬细胞在相同的AgNP浓度水平和暴露时间下对AgNP的摄取，这表明依赖于暴露浓度和更高的AgNP摄取量（每个细胞大约9至45个AgNP）要高得多。随时间保持不变。通过样品消解，ICP-MS分析以及其他提供单细胞分析的有前途的替代技术，对结果进行了比较和验证。

更新日期：2018-05-18

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