TY - JOUR A1 - Drescher, Daniela A1 - Giesen, Charlotte A1 - Traub, Heike A1 - Panne, Ulrich A1 - Kneipp, Janina A1 - Jakubowski, Norbert T1 - Quantitative imaging of gold and silver nanoparticles in single eukaryotic cells by laser ablation ICP-MS N2 - Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was utilized for spatially resolved bioimaging of the distribution of silver and gold nanoparticles in individual fibroblast cells upon different incubation experiments. High spatial resolution was achieved by optimization of scan speed, ablation frequency, and laser energy. Nanoparticles are visualized with respect to cellular substructures and are found to accumulate in the perinuclear region with increasing incubation time. On the basis of matrix-matched calibration, we developed a method for quantification of the number of metal nanoparticles at the single-cell level. The results provide insight into nanoparticle/cell interactions and have implications for the development of analytical methods in tissue diagnostics and therapeutics. KW - Imaging KW - Cell KW - Nanoparticles KW - Laser ablation KW - ICP-MS PY - 2012 U6 - https://doi.org/10.1021/ac302639c SN - 0003-2700 SN - 1520-6882 VL - 84 IS - 22 SP - 9684 EP - 9688 PB - American Chemical Society CY - Washington, DC AN - OPUS4-27440 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CHAP A1 - Theiner, S. A1 - Corte Rodriguez, M. A1 - Traub, Heike ED - Golloch, A. T1 - Novel applications of lanthanoids as analytical or diagnostic tools in the life sciences by ICP-MS based techniques N2 - Inductively coupled plasma-mass spectrometry (ICP-MS) is a well-established analytical method offering high sensitivity and multi-element analysis. ICP-MS has found acceptance in various application areas ranging from material analysis to applications in the life sciences. Within the last 15 years new strategies for the sensitive detection and accurate quantification of biomolecules in complex biomedical samples have been developed. Recent instrumental improvements have contributed to this progress. As most of the biomolecules do not contain endogenous metals etectable with ICP-MS, bioconjugation with artificial metal-containing tags based on metal-loaded chelate complexes or nanoparticles is increasingly applied to determine biomolecules indirectly. Especially, the combination of immunohistochemical workflows using lanthanoid-tagged antibodies and ICP-MS detection provides new insights in the complexity and interdependency of cellular processes. Single-cell ICP-MS, also termed as mass cytometry, allows high-dimensional analysis of biomarkers in cell populations at single-cell resolution. For that purpose, lanthanoid isotope labelled antibodies are used to detect their corresponding target molecules. The visualisation of the elemental distribution is possible with laser ablation ICP-MS (LA-ICPMS) at high spatial resolution. Especially, the combination of LA with ICP time-of-flight mass spectrometry, also referred to as imaging mass cytometry (IMC), opens new possibilities for multiparametric tissue imaging at the single-cell level and even below. The lanthanoid localisation and concentration can be linked to their conjugated antibody target providing valuable information about surface markers, intracellular signalling molecules to measure biological function, and the network state of an individual cell in a tissue. This book chapter focuses on new applications, where the multi-element capabilities of ICP-MS are used for the detection of lanthanoids applied as artificial elemental stains or tags for biomolecules and in particular antibodies. KW - ICP-MS KW - Laser ablation KW - Cell KW - Antibody KW - Immunohistochemistry KW - Lanthanoid KW - Mass cytometry KW - Imaging PY - 2022 SN - 978-3-11069-645-5 SN - 978-3-11069-636-3 U6 - https://doi.org/10.1515/9783110696455-013 SP - 399 EP - 444 PB - De Gruyter CY - Berlin, Boston ET - 2. rev. and exten. edition AN - OPUS4-55118 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CHAP A1 - Traub, Heike ED - Milacic, R. ED - Scancar, J. ED - Goenaga-Infante, H. ED - Vidmar, J. T1 - Imaging of metal-based nanoparticles in tissue and cell samples by laser ablation inductively coupled plasma mass spectrometry N2 - Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is nowadays a versatile and powerful analytical method for direct solid sample analysis. The applicability has been demonstrated for a wide variety of samples covering hard and soft materials. In an imaging mode the technique provides quantitative information on the elemental distribution within a sample. LA-ICP-MS imaging is of particular interest in biomedical research as the distribution of an element gives valuable insight on uptake and distribution of essential and toxic trace elements, administered contrast agents as well es nanoparticles. LA-ICP-MS is therefore a powerful complement to other imaging techniques. Recent instrumental improvements, especially in sample chamber design, have contributed to better sensitivity and spatial resolution enabling subcellular imaging. The book chapter provides a comprehensive overview about spatially resolved localisation and quantification of various nanoparticles in cells and tissue thin sections by LA-ICP-MS. Furthermore, different sample preparation strategies and internal standardisation and calibration approaches for bioimaging by LA-ICP-MS are summarized and discussed. Metal-containing nanomaterials are used in numerous fields ranging from industrial applications to nanomedicine. Several studies have demonstrated that the physicochemical properties of nanoparticles have an impact on their pharmacokinetics, transfer and clearance. The high sensitivity and multielement capability of LA-ICP-MS enables the elucidation of interactions between tissue components and nanomaterials used as imaging probes or drug carriers. Potential toxic effects are investigated as well. Thus, LA imaging significantly supports the clinical translation of safe and efficient nanoparticles for diagnostic and therapeutic purposes. KW - Laser ablation KW - ICP-MS KW - Imaging KW - Nanoparticle KW - Nanomaterial KW - Tissue KW - Cell PY - 2021 SN - 978-0-323-85305-7 SN - 0166-526X VL - 93 SP - 173 EP - 240 PB - Elsevier CY - Amsterdam ET - 1 AN - OPUS4-52775 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Traub, Heike A1 - Drescher, Daniela A1 - Büchner, T. A1 - Zeise, Ingrid A1 - Kneipp, Janina A1 - Jakubowski, Norbert T1 - Studying cellular uptake and processing of nanoparticles by LA-ICP-MS N2 - In recent years, elemental imaging of biological samples using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is gaining in importance. Latest improvements regarding spatial resolution (down to 1 µm) and washout time make LA-ICP-MS particularly interesting for single cell analysis. Many current nanomaterials can serve as contrast agents in cellular or tissue imaging, drug delivery vehicles or therapeutics, whereas others can cause toxic effects. In order to evaluate nano-bio interactions, the number of nanoparticles (NPs) inside cells as well as their localisation within cellular substructures is of particular interest. LA-ICP-MS was used to study the NP pathway from uptake, via intracellular processing up to cell division. Fibroblast cells were incubated with different metallic NPs under varying experimental conditions. For LA analysis the cells were fixed with formaldehyde and dried. Our results show that LA-ICP-MS is able to localise NP aggregates within cellular substructures. The NPs accumulate in the perinuclear region in the course of intracellular processing, e.g. multivesicular fusion and endosomal maturation, but do not enter the nucleus [1, 2]. A strong dependence of NP uptake on concentration and incubation time was found. Additionally, the number of NPs internalized by individual cells was determined and variations within the cell population became visible. A new laser ablation system providing a short washout time (50 ms) together with small spot sizes (< 4 µm) and high repetition rates allows high spatial resolution applications. First results of cell imaging will be shown. The findings demonstrate the potential of LA-ICP-MS enabling insight into NP uptake and intracellular distribution dependent on experimental parameters. T2 - 8th Nordic Conference on Plasma Spectrochemistry CY - Loen, Norway DA - 05.06.2016 KW - Imaging KW - LA-ICP-MS KW - Cell KW - Nanoparticles PY - 2016 AN - OPUS4-36500 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Büchner, T. A1 - Drescher, D. A1 - Merk, V. A1 - Traub, Heike A1 - Guttmann, P. A1 - Werner, St. A1 - Jakubowski, Norbert A1 - Schneider, G. A1 - Kneipp, J. T1 - Biomolecular environment, quantification, and intracellular interaction of multifunctional magnetic SERS nanoprobes N2 - Multifunctional composite nanoprobes consisting of iron oxide nanoparticles linked to silver and gold nanoparticles, Ag–Magnetite and Au–Magnetite, respectively, were introduced by endocytic uptake into cultured fibroblast cells. The cells containing the non-toxic nanoprobes were shown to be displaceable in an external magnetic field and can be manipulated in microfluidic channels. The distribution of the composite nanostructures that are contained in the endosomal system is discussed on the basis of surfaceenhanced Raman scattering (SERS) mapping, quantitative laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) micromapping, and cryo soft X-ray tomography (cryo soft-XRT). Cryo soft-XRT of intact, vitrified cells reveals that the composite nanoprobes form intra-endosomal aggregates. The nanoprobes provide SERS signals from the biomolecular composition of their surface in the endosomal environment. The SERS data indicate the high stability of the nanoprobes and of their plasmonic properties in the harsh environment of endosomes and lysosomes. The spectra point at the molecular composition at the surface of the Ag–Magnetite and Au–Magnetite nanostructures that is very similar to that of other Composite structures, but different from the composition of pure silver and gold SERS nanoprobes used for intracellular investigations. As shown by the LA-ICP-MS data, the uptake efficiency of the magnetite composites is approximately two to three times higher than that of the pure gold and silver nanoparticles. KW - Nanoparticles KW - SERS KW - Cell KW - LA-ICP-MS KW - X-ray tomography PY - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-371811 SN - 0003-2654 VL - 141 IS - 17 SP - 5096 EP - 5106 PB - Royal Society of Chemistry CY - Cambridge, UK AN - OPUS4-37181 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Traub, Heike A1 - Drescher, Daniela A1 - Büchner, T. A1 - Kneipp, Janina A1 - Jakubowski, Norbert T1 - Studying cellular uptake and processing of nanoparticles by LA-ICP-MS N2 - In recent years, elemental imaging of biological samples using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is gaining in importance. Recent improvements regarding spatial resolution (down to 1 µm) and washout time make LA-ICP-MS particularly interesting for single cell analysis. Many current nanomaterials can serve as contrast agents in cellular or tissue imaging, drug delivery vehicles or therapeutics, whereas others can cause toxic effects. In order to evaluate nano-bio interactions, the number of nanoparticles (NPs) inside cells as well as their localisation within cellular substructures is of particular interest. LA-ICP-MS was used to study the NP pathway from uptake, via intracellular processing up to cell division. Fibroblast cells were incubated with different metallic NPs under varying experimental conditions. For LA analysis the cells were fixed with formaldehyde and dried. Our results show that LA-ICP-MS is able to localise NP aggregates within cellular substructures. The NPs accumulate in the perinuclear region in the course of intracellular processing, e.g. multivesicular fusion and endosomal maturation, but do not enter the nucleus [1, 2]. A strong dependence of NP uptake on concentration and incubation time was found. Additionally, the number of NPs internalized by individual cells was determined and variations within the cell population became visible. A new laser ablation system providing a short washout time (50 ms) together with small spot sizes (< 4 µm) and high repetition rates allows high spatial resolution applications. First results of cell imaging will be shown. The findings demonstrate the potential of LA-ICP-MS providing insight into NP uptake and intracellular distribution dependent on experimental parameters. T2 - 13th European Workshop on Laser Ablation CY - Ljubljana, Slovenia DA - 12.07.2016 KW - LA-ICP-MS KW - Nanoparticle KW - Bio-imaging KW - Cell PY - 2016 AN - OPUS4-36891 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Büchner, Tina A1 - Drescher, Daniela A1 - Traub, Heike A1 - Schrade, P. A1 - Bachmann, S. A1 - Jakubowski, Norbert A1 - Kneipp, Janina T1 - Relating surface-enhanced Raman scattering signals of cells to gold nanoparticle aggregation as determined by LA-ICP-MS micromapping N2 - The cellular response to nanoparticle exposure is essential in various contexts, especially in nanotoxicity and nanomedicine. Here, 14-nm gold nanoparticles in 3T3 fibroblast cells are investigated in a series of pulse-chase experiments with a 30-min incubation pulse and chase times ranging from 15 min to 48 h. The gold nanoparticles and their aggregates are quantified inside the cellular ultrastructure by laser ablation inductively coupled plasma mass spectrometry micromapping and evaluated regarding the surface-enhanced Raman scattering (SERS) signals. In this way, both information about their localization at the micrometre scale and their molecular nanoenvironment, respectively, is obtained and can be related. Thus, the nanoparticle pathway from endocytotic uptake, intracellular processing, to cell division can be followed. It is shown that the ability of the intracellular nanoparticles and their accumulations and aggregates to support high SERS signals is neither directly related to nanoparticle amount nor to high local nanoparticle densities. The SERS data indicate that aggregate geometry and interparticle distances in the cell must change in the course of endosomal maturation and play a critical role for a specific gold nanoparticle type in order to act as efficient SERS nanoprobe. This finding is supported by TEM images, showing only a minor portion of aggregates that present small interparticle spacing. The SERS spectra obtained after different chase times show a changing composition and/or structure of the biomolecule corona of the gold nanoparticles as a consequence of endosomal processing. KW - Gold nanoparticles KW - Surface-enhanced Raman scattering KW - LA-ICP-MS KW - Fibroblast KW - Cell KW - Particle aggregation KW - Endosome PY - 2014 U6 - https://doi.org/10.1007/s00216-014-8069-0 SN - 1618-2642 SN - 1618-2650 VL - 406 IS - 27 SP - 7003 EP - 7014 PB - Springer CY - Berlin AN - OPUS4-31718 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Traub, Heike A1 - Drescher, D. A1 - Büchner, T. A1 - Merk, V. A1 - Kneipp, J. A1 - Jakubowski, Norbert T1 - Studying nanoparticle-cell interaction by LA ICP-MS N2 - The interaction of nanoparticles (NPs) with cells has become a major field of interest, ranging from medical applications to nanotoxicology. Size, shape and surface modification of the NPs determine the uptake rate and pathway into the cells, and therefore impact specific cell components and processes. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is an established quantitative multi-elemental analysis and mapping technique. In recent years, it was shown that LA-ICP-MS can provide quantitative as well as distribution information of NPs in cell samples. Here LA-ICP-MS was applied for the imaging of individual cells to study the uptake and intracellular processing of metal-containing nanostructures. Additionally, the local distribution of naturally occurring elements in cells like P was measured to indicate cell morphology. The cells were incubated with different types of NPs under varying experimental conditions. For LA analysis, the cells were fixed and dried. Our findings show, that LA-ICP-MS is suitable for the localisation of nanoparticle aggregates within cellular compartments. The studied NPs accumulate in the perinuclear region in the course of intracellular processing, but do not enter the cell nucleus. The uptake efficiency depends strongly on the physicochemical properties of the nanostructures as well as on the incubation conditions like concentration and incubation time. The results demonstrate the potential of LA-ICP-MS providing insight into NP uptake, intracellular distribution and cell-to-cell variation dependent on experimental parameters. T2 - Workshop on Laser Bioimaging Mass Spectrometry CY - Münster, Germany DA - 24.05.2018 KW - Imaging KW - Laser ablation KW - ICP-MS KW - Nanoparticle KW - Cell PY - 2018 AN - OPUS4-45071 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pisonero, J. A1 - Bouzas-Ramos, D. A1 - Traub, Heike A1 - Cappella, Brunero A1 - Alvarez-Llamas, C. A1 - Richter, Silke A1 - Mayo, J. C. A1 - Costa-Fernandez, J. M. A1 - Bordel, N. A1 - Jakubowski, Norbert T1 - Critical evaluation of fast and highly resolved elemental distribution in single cells using LA-ICP-SFMS N2 - The analytical potential of a nanosecond laser ablation inductively coupled plasma mass spectrometer (ns-LA-ICP-SFMS) system, equipped with an ultra-fast wash-out ablation chamber, is critically investigated for fast and highly spatially resolved (∼μm) qualitative elemental distribution within single cells. Initially, a low surface roughness (< 10 nm) thin In–SnO2 layer (total coating thickness ∼200 nm) deposited on glass is employed to investigate the size, morphology and overlapping of laser-induced craters obtained at different laser repetition rates, making use of Atomic Force Microscopy (AFM). Conical craters with a surface diameter of about 2 µm and depths of about 100 nm were measured after a single laser shot. Furthermore, the influence of the sampling distance (i.e. distance between the sample surface and the inner sniffer of the ablation chamber) on the LA-ICP-MS ion signal wash-out time is evaluated. A significant decrease of the transient 120Sn+ ion signal is noticed after slight variations (±200 μm) around the optimum sampling position. Ultra-fast wash-outs (< 10 ms) are achieved reducing the aerosol mixing from consecutive laser shots even when operating the laser at high repetition rates (25 – 100 Hz). Fast and highly spatially resolved images of elemental distribution within mouse embryonic fibroblast cells (NIH/3T3 fibroblast cells) and human cervical carcinoma cells (HeLa cells), incubated with gold nanoparticles (Au NPs) and Cd-based quantum dots (QDs), respectively, are determined at the optimized operating conditions. Elemental distribution of Au and Cd in single cells is achieved using a high scanning speed (50 µm/s) and high repetition rate (100 Hz). The results obtained for the distribution of fluorescent Cd-based QDs within the HeLa cells are in good agreement with those obtained by confocal microscopy. The size, morphology and overlapping of laser-induced craters in the fixed cells are also investigated using AFM, observing conical craters with a surface diameter of about 2.5 µm and depths of about 800 nm after a single laser shot. KW - Laser ablation KW - ICP-MS KW - Nanoparticle KW - Atomic Force Microscopy KW - Cell PY - 2018 U6 - https://doi.org/10.1039/c8ja00096d SP - 1 EP - 9 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-45564 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Traub, Heike A1 - Drescher, D. A1 - Büchner, T. A1 - Pisonero, J. A1 - Bouzas-Ramos, D. A1 - Kneipp, J. A1 - Jakubowski, Norbert T1 - LA-ICP-MS to study nanoparticle-cell interaction N2 - Nanoparticles (NPs) have found a wide range of applications in research and industry. Thereby the interaction of NPs with biological systems like cells has become a major field of interest, ranging from medical applications to nanotoxicology. Size, shape and surface modification of the nanomaterials determine the uptake rate and pathway into the cells, and therefore impact specific cell components and processes. In recent years, elemental imaging of biological samples using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is gaining more and more importance. Improvements concerning both spatial resolution (down to 1 µm) and signal-to-background ratio due to low-dispersion LA chambers make LA-ICP-MS particularly interesting for single cell analysis. Here LA-ICP-MS was applied for the imaging of individual cells to study the uptake and intracellular processing of metal-containing nanostructures. The cells were incubated with different NPs under varying experimental conditions and afterwards fixed with para-formaldehyde and dried for LA analysis. High-spatial resolution LA-ICP-MS was achieved by careful optimisation of the laser ablation parameters. Our findings show, that LA-ICP-MS is applicable to localize NP aggregates within cellular compartments. The uptake efficiency depends strongly on the physicochemical properties of the nanostructures as well as on the incubation conditions like concentration and incubation time. The results demonstrate the potential of LA-ICP-MS providing insight into nanoparticle-cell interaction dependent on experimental parameters. T2 - 14th European Workshop on Laser Ablation (EWLA) CY - Pau, France DA - 26.06.2018 KW - Laser ablation KW - ICP-MS KW - Imaging KW - Nanoparticle KW - Cell PY - 2018 AN - OPUS4-45570 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -