Quantification of silver nanoparticles taken up by single cells using inductively coupled plasma mass spectrometry in the single cell measurement mode

Oliver, Ana López-Serrano; Baumgart, Sabine; Bremser, Wolfram; Flemig, Sabine; Wittke, Doreen; Grützkau, Andreas; Luch, Andreas; Haase, Andrea; Jakubowski, Norbert

doi:10.1039/c7ja00395a

Cited by 36 publications

(17 citation statements)

References 19 publications

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“…The absolute amount of AgNPs per cell was time- and dose-dependent and ranged from 4 to 75, i.e., 3–56 fg Ag/cell. These results are in accordance with those ones reported for less maturated, nonadherent THP-1 cells measured by SC-ICPMS …”

Section: Resultssupporting

confidence: 93%

“…from DNA staining or from the NP metal core. Here, the transient ion signal is integrated over a single event. , To quantify the AgNP uptake of THP-1 cells at the single cell level, a similar external calibration as previously described was performed by measuring AgNP suspensions with increasing Ag concentrations. As indicated in Table S-2, proportional to the AgNP concentration, the number of particle events exhibits a linear increase ( R 2 = 0.99615).…”

Section: Resultsmentioning

confidence: 99%

“…Recently, the slice and view technology of fast ion bombardment (FIB) combined with the secondary electron microscopy (SEM) technique was used to quantify 75 nm silver NPs (AgNPs) in a single cell but has limited high-throughput capacities . Inductively coupled plasma mass spectrometry (ICPMS), traditionally a bulk measurement of lysed NP-exposed cells that provides averaged data, , but operated in “single particle” detection mode enables this methodology to enter the field of single-cell analysis (SC-ICPMS). , Although ICPMS covers a wide mass range from 7 to 250 atomic mass units (amu), SC-ICPMS with a quadrupole or sector-field mass analyzer is limited to record only one isotope in the short duration of a cell event . Therefore, SC-ICPMS alone does not allow multiparametric analyses at the single cell level, which are usually necessary in many fields of basic and medical research.…”

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confidence: 99%

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Mass Cytometry Enabling Absolute and Fast Quantification of Silver Nanoparticle Uptake at the Single Cell Level

et al. 2019

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In the last decades, significant efforts have been made to investigate possible cytotoxic effects of metallic nanoparticles (NPs). Methodologies enabling precise information regarding uptake and intracellular distribution of NPs at the single cell level remain to be established. Mass cytometry (MC) has been developed for high-dimensional single cell analyses and is a promising tool to quantify NP−cell interactions. Here, we aim to establish a new MC-based quantification procedure to receive absolute numbers of NPs per single cell by using a calibration that considers the specific transmission efficiency (TE) of suspended NPs. The current MC-quantification strategy accept TE values of complementary metal solutions. In this study, we demonstrate the different transmission behavior of 50 nm silver NPs (AgNP) and silver nitrate solution. We have used identical AgNPs for calibration as for in vitro-differentiated macrophages (THP-1 cell line) in a time-and dose-dependent manner. Our quantification relies on silver intensities measuring AgNPs in the same detection mode as the cells. Results were comparable with the TE quantification strategy using AgNPs but differed when using ionic silver. Furthermore, intact and digested cell aliquots were measured to investigate the impact of MC sample processing on the amount of AgNPs/cell. Taken together, we have provided a MC-specific calibration procedure to precisely calculate absolute numbers of NPs per single cell. Combined with its unique feature of multiplexing up to 50 parameters, MC provides much more information on the single cell level than single cell-inductively coupled plasma mass spectrometry (SC-ICPMS) and, therefore, offers new opportunities in nanotoxicology.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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Mass Cytometry Enabling Absolute and Fast Quantification of Silver Nanoparticle Uptake at the Single Cell Level

et al. 2019

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“…The validation was based on ∼1000 cells for laser ablation and ∼20 000 cells for solution-based analysis. As a result of the integration time of 3 ms in solution measurements, most likely double cell events could not be excluded, 5 explaining the observed cell events with higher platinum levels and thus larger distribution as compared to bioimaging experiments.…”

Section: Results and Discussionmentioning

confidence: 97%

Cisplatin Uptake in Macrophage Subtypes at the Single-Cell Level by LA-ICP-TOFMS Imaging

et al. 2021

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A high-throughput laser ablation–inductively coupled plasma–time-of-flight mass spectrometry (LA-ICP-TOFMS) workflow was implemented for quantitative single-cell analysis following cytospin preparation of cells. For the first time, in vitro studies on cisplatin exposure addressed human monocytes and monocyte-derived macrophages (undifferentiated THP-1 monocytic cells, differentiated M0 macrophages, as well as further polarized M1 and M2 phenotypes) at the single-cell level. The models are of particular interest as macrophages comprise the biggest part of immune cells present in the tumor microenvironment and play an important role in modulating tumor growth and progression. The introduced bioimaging workflow proved to be universally applicable to adherent and suspension cell cultures and fit-for-purpose for the quantitative analysis of several hundreds of cells within minutes. Both, cross-validation of the method with single-cell analysis in suspension for THP-1 cells and with LA-ICP-TOFMS analysis of adherent M0 cells grown on chambered glass coverslips, revealed agreeing platinum concentrations at the single-cell level. A high incorporation of cisplatin was observed in M2 macrophages compared to the M0 and M1 macrophage subtypes and the monocyte model, THP-1. The combination with bright-field images and monitoring of highly abundant endogenous elements such as phosphorus and sodium at a high spatial resolution allowed assessing cell size and important morphological cell parameters and thus straightforward control over several cell conditions. This way, apoptotic cells and cell debris as well as doublets or cell clusters could be easily excluded prior to data evaluation without additional staining.

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“…In Table , only results for Ag are shown as the concentrations for all other trace and mineral elements measured after digestion have been below the limits of detection of liquid ICP-SF-MS analysis. This was because LODs were significantly lower for LA-ICP-MS due to the following facts: first, a dilution of sample by digestion is avoided in direct measurements by LA-ICP-MS; second, there is no contamination from solvent; and third, the transport efficiency of LA-ICP-MS is close to 100%, whereas much lower efficiencies of 5% at most have been reported for pneumatic nebulization . To determine other elements within the MCS, more than a few MCSs must be digested even in the case of phosphorus, which has the highest content of elements quantified.…”

Section: Resultsmentioning

confidence: 99%

Quantitative Imaging of Silver Nanoparticles and Essential Elements in Thin Sections of Fibroblast Multicellular Spheroids by High Resolution Laser Ablation Inductively Coupled Plasma Time-of-Flight Mass Spectrometry

et al. 2019

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We applied high resolution laser ablation inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-TOF-MS) with cellular spatial resolution for bioimaging of nanoparticles uptaken by fibroblast multicellular spheroids (MCS). This was used to quantitatively investigate interactions of silver nanoparticles (Ag NPs) and the distributions of intrinsic minerals and biologically relevant elements within thin sections of a fibroblast MCS as a three-dimensional in vitro tissue model. We designed matrix-matched calibration standards for this purpose and printed them using a noncontact piezo-driven array spotter with a Ag NP suspension and multielement standards. The limits of detection for Ag, Mg, P, K, Mn, Fe, Co, Cu, and Zn were at the femtogram (10–15 g) level, which is sufficient to investigate intrinsic minerals in thin MCS sections (20 μm thick). After incubation for 48 h, Ag NPs were enriched in the outer rim of the MCS but not detected in the core. The localization of Ag NPs was inhomogeneous in the outer rim, and they were colocalized with a single-cell-like structure visualized by Fe distribution (pixel size of elemental images: 5 × 0.5 μm). The quantitative value for the total mass of Ag NPs in a thin section by the present method agreed with that obtained by ICP-sector field (SF)-MS with a liquid mode after acid digestion.

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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

Abstract: A promising analytical methodology is proposed to study nanoparticle-cell interactions providing information of the number of NPs internalized by cells or externally bound to the cell surface.

Cited by 36 publications

References 19 publications

Mass Cytometry Enabling Absolute and Fast Quantification of Silver Nanoparticle Uptake at the Single Cell Level

Mass Cytometry Enabling Absolute and Fast Quantification of Silver Nanoparticle Uptake at the Single Cell Level

Cisplatin Uptake in Macrophage Subtypes at the Single-Cell Level by LA-ICP-TOFMS Imaging

Quantitative Imaging of Silver Nanoparticles and Essential Elements in Thin Sections of Fibroblast Multicellular Spheroids by High Resolution Laser Ablation Inductively Coupled Plasma Time-of-Flight Mass Spectrometry

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