Stijn Van Malderen

Development of a fast laser ablation-inductively coupled plasma-mass spectrometry cell for sub-μm scanning of layered materials

Performance data are reported for a commercially available laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) setup, equipped with a custom-made cell. The low dispersion ablation cell and the connecting tubing achieve a 99% washout of the aerosol in ∼6 ms, enabling separated pulse responses at frequencies up to 200–300 Hz. In addition, the cell employed supports a post-acquisition methodology for the deconvolution of overlapping ablation positions in scanning mode by an iterative Richardson–Lucy algorithm. This enables correction of the distortion in the scan profile upon traversing layers with dimensions below the physical size of the laser beam. By overlapping the ablation positions of a 1 μm diameter laser beam, a lateral resolution in the order of 0.3 ± 0.1 μm was demonstrated for scanning of μm-sized layers in high capacitance multi-layer ceramic capacitors.

Recent developments in the design of rapid response cells for laser ablation-inductively coupled plasma-mass spectrometry and their impact on bioimaging applications

This review covers developments in the design of Laser Ablation (LA) cells, the associated transport tubing assembly, and their coupling to Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) instrumentation. Recent ablation cell designs have reduced the pulse response duration for a single laser shot to <10 ms, using the criterion of the full peak width at 1% of the height of the maximum signal intensity. The evolution towards these low dispersion systems has been profoundly influenced by our understanding of processes driving the initial dispersion, of the design aspects of the cell and tubing that influence transport-induced dispersion and transport efficiency, and of limitations imposed by the temporal resolution of ICP-MS instruments, all of which are discussed. Rapid response LA-ICP-MS systems greatly benefit throughput and sensitivity, which are key parameters in 2D and 3D imaging at high lateral resolution. The analysis and imaging of biological material has come to the forefront as a key application of LA-ICP-MS. The impact of the technical developments in LA-ICP-MS systems on emerging applications, including multiplexed metal-tagged antibody detection (for immunohistochemistry), nanoparticle and compound hypo- and hyperaccumulation, and (intra-) cellular/histological studies, is also discussed.

Submicrometer Imaging by Laser Ablation-Inductively Coupled Plasma Mass Spectrometry via Signal and Image Deconvolution Approaches

In this work, pre- and postacquisition procedures for enhancing the lateral resolution of laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS) in two- and three-dimensional (2D, 3D) nuclide distribution mapping beyond the laser beam waist are described. 2D images were constructed by projecting a rectangular grid of discrete LA positions, arranged at interspacings smaller than the dimensions of the laser beam waist, onto the sample surface, thus oversampling the region of interest and producing a 2D image convolved in the spatial domain. The pulse response peaks of a low-dispersion LA cell were isolated via signal deconvolution of the transient mass analyzer response. A 3D stack of 2D images was deconvolved by an iterative Richardson-Lucy algorithm with Total Variance regularization, enabling submicrometer image fidelity, demonstrated in the analysis of trace level features in corroded glass. A point spread function (PSF) could be derived from topography maps of single pulse craters from atomic force microscopy. This experimental PSF allows the approach to take into account the laser beam shape, beam aberrations, and the laser-solid interaction, which in turn enhances the spatial resolution of the reconstructed volume.

High-resolution laser ablation-inductively coupled plasma-mass spectrometry imaging of cisplatin-induced nephrotoxic side effects

Two-dimensional elemental mapping (bioimaging) via laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was performed on 5xa0μm thick formalin-fixed, paraffin-embedded kidney tissue sections from Cynomolgus monkeys administered with increasing pharmacological doses of cisplatin. Laterally resolved pixels of 1xa0μm were achieved, enabling elemental analysis on a (sub-)cellular level. Zones of high Pt response were observed in the renal cortex, where proximal tubules are present, the epithelium of which is responsible for partial reabsorption of cisplatin. Histopathological evaluation, of hematoxylin and eosin-stained serial sections, adjacent to the sections probed via LA-ICP-MS, revealed minimal to mild cisplatin-related lesions (<100xa0μm) in the renal cortex. Necrotic proximal tubules with sloughed epithelial cells in their lumen could be linked directly to the areas with the highest accumulation of cisplatin, indicating a direct link between cellular concentration and toxicity, thereby providing more insight into the mechanisms through which renal damage occurs.

Imaging the 3D trace metal and metalloid distribution in mature wheat and rye grains via laser ablation-ICP-mass spectrometry and micro-X-ray fluorescence spectrometry

Toxic trace metals and metalloids in human nutrient sources pose a severe health risk, and the processes governing metal accumulation should hence be well understood. In this work, the spatial distribution of toxic trace metals/metalloids and micronutrients (Cr, Mn, Ni, Cu, Zn, As, Cd, Hg and Pb) in mature wheat (Triticum aestivum L.) and rye (Secale cereale L.) grains at typical exposure levels was visualized and quantified via laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) by serial sectioning. The bulk concentrations of these elements were also determined by pneumatic nebulisation-ICP-MS. Furthermore, longitudinal sections were scanned using m-X-ray fluorescence spectrometry to confirm the major element distribution. Serial sectioning in this study was realized via a polishing strategy. Although the methodology is time-consuming and laborious, it enables to access 3D information for samples which cannot be sectioned using a microtome on a depth scale that would otherwise be inaccessible by a laser probe. In the elemental images, strong local enrichment patterns for Mn and Zn are apparent in the aleurone layer/seed coat, vascular tissue of the crease, and embryonic tissue, whereas Cr, As, Cd and Pb have been mainly accumulated in the grain endosperm as a result of different transport and storage dynamics.

Three-Dimensional Reconstruction of the Tissue-Specific Multielemental Distribution within Ceriodaphnia dubia via Multimodal Registration Using Laser Ablation ICP-Mass Spectrometry and X-ray Spectroscopic Techniques

In this work, the three-dimensional elemental distribution profile within the freshwater crustacean Ceriodaphnia dubia was constructed at a spatial resolution down to 5 μm via a data fusion approach employing state-of-the-art laser ablation-inductively coupled plasma-time-of-flight mass spectrometry (LA-ICP-TOFMS) and laboratory-based absorption microcomputed tomography (μ-CT). C. dubia was exposed to elevated Cu, Ni, and Zn concentrations, chemically fixed, dehydrated, stained, and embedded, prior to μ-CT analysis. Subsequently, the sample was cut into 5 μm thin sections that were subjected to LA-ICP-TOFMS imaging. Multimodal image registration was performed to spatially align the 2D LA-ICP-TOFMS images relative to the corresponding slices of the 3D μ-CT reconstruction. Mass channels corresponding to the isotopes of a single element were merged to improve the signal-to-noise ratios within the elemental images. In order to aid the visual interpretation of the data, LA-ICP-TOFMS data were projected onto the μ-CT voxels representing tissue. Additionally, the image resolution and elemental sensitivity were compared to those obtained with synchrotron radiation based 3D confocal μ-X-ray fluorescence imaging upon a chemically fixed and air-dried C. dubia specimen.

Hybrid imaging labels : Providing the link between mass spectrometry-based molecular pathology and theranostics

Background: Development of theranostic concepts that include inductively coupled plasma mass spectrometry (ICP-MS) and laser ablation ICP-MS (LA-ICP-MS) imaging can be hindered by the lack of a direct comparison to more standardly used methods for in vitro and in vivo evaluation; e.g. fluorescence or nuclear medicine. In this study a bimodal (or rather, hybrid) tracer that contains both a fluorescent dye and a chelate was used to evaluate the existence of a direct link between mass spectrometry (MS) and in vitro and in vivo molecular imaging findings using fluorescence and radioisotopes. At the same time, the hybrid label was used to determine whether the use of a single isotope label would allow for MS-based diagnostics. Methods: A hybrid label that contained both a DTPA chelate (that was coordinated with either 165Ho or 111In) and a Cy5 fluorescent dye was coupled to the chemokine receptor 4 (CXCR4) targeting peptide Ac-TZ14011 (hybrid-Cy5-Ac-TZ4011). This receptor targeting tracer was used to 1) validate the efficacy of (165Ho-based) mass-cytometry in determining the receptor affinity via comparison with fluorescence-based flow cytometry (Cy5), 2) evaluate the microscopic binding pattern of the tracer in tumor cells using both fluorescence confocal imaging (Cy5) and LA-ICP-MS-imaging (165Ho), 3) compare in vivo biodistribution patterns obtained with ICP-MS (165Ho) and radiodetection (111In) after intravenous administration of hybrid-Cy5-Ac-TZ4011 in tumor-bearing mice. Finally, LA-ICP-MS-imaging (165Ho) was linked to fluorescence-based analysis of excised tissue samples (Cy5). Results: Analysis with both mass-cytometry and flow cytometry revealed a similar receptor affinity, respectively 352 ± 141 nM and 245 ± 65 nM (p = 0.08), but with a much lower detection sensitivity for the first modality. In vitro LA-ICP-MS imaging (165Ho) enabled clear discrimination between CXCR4 positive and negative cells, but fluorescence microscopy was required to determine the intracellular distribution. In vivo biodistribution patterns obtained with ICP-MS (165Ho) and radiodetection (111In) of the hybrid peptide were shown to be similar. Assessment of tracer distribution in excised tissues revealed the location of tracer uptake with both LA-ICP-MS-imaging and fluorescence imaging. Conclusion: Lanthanide-isotope chelation expands the scope of fluorescent/radioactive hybrid tracers to include MS-based analytical tools such as mass-cytometry, ICP-MS and LA-ICP-MS imaging in molecular pathology. In contradiction to common expectations, MS detection using a single chelate imaging agent was shown to be feasible, enabling a direct link between nuclear medicine-based imaging and theranostic methods.

Fast High-Resolution Laser Ablation-Inductively Coupled Plasma Mass Spectrometry Imaging of the Distribution of Platinum-Based Anticancer Compounds in Multicellular Tumor Spheroids

Multicellular tumor spheroid models serve as an important three-dimensional in vitro cell model system as they mimic the complex tumor microenvironment and thus have contributed to valuable assays in drug discovery studies. In this study, we present a state-of-the-art laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS) setup for high spatial resolution elemental imaging of multicellular tumor spheroids and an approach to account for variations in cell density. A low dispersion LA-ICPMS setup was employed, providing accelerated throughput and high sensitivity and permitting a lateral image resolution down to ∼2.5 μm for phosphorus and platinum in HCT116 colon cancer spheroids upon treatment with the clinically used anticancer drug oxaliplatin. Phosphorus was introduced as scalar to compensate for differences in cell density and tissue thickness and the Pt/P ratios together with the high resolution adopted in our approach allows the differentiation of platinum accumulation within each part of the morphology of the tumor spheroids (layers of proliferating, quiescent, and necrotic cells).

Imaging artifacts in continuous scanning 2D LA-ICPMS imaging due to nonsynchronization issues

Pulsed laser ablation (LA) devices in laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) imaging have become very advanced, delivering laser pulses with high temporal accuracy and stable energy density. However, unintentional imaging artifacts may be generated in 2D element maps when the LA repetition rate and the data acquisition parameters of ICPMS instruments with a sequential mass spectrometer (i.e., quadrupole filter or sector-field mass spectrometer) are desynchronized. This may potentially lead to interference patterns, visible as ripples in elemental images, and thus, compromised image quality. This paper describes the background of aliasing in continuous scanning mode through simulation experiments and ways to modulate the effect. The existence of this image degradation source is demonstrated experimentally via real-life imaging of a homogeneous glass standard.

High-speed sub-micrometer imaging of sub-cellular structures in single cells using ARIS

© 2017 Teledyne CETAC Technologies TECHNICAL NOTE: ANALYTE G2-003 High-speed sub-micrometer imaging of sub-cellular structures in single cells using ARIS Thibaut Van Acker1, Stijn J.M. Van Malderen1, Tessa Buckle2, Frank Vanhaecke1 1Department of Analytical Chemistry, Ghent University, Campus Sterre, Krijgslaan 281 S12, 9000 Ghent, Belgium 2Interventional Molecular Imaging Laboratory, Department of Radiology, Leiden University Medical Center, Albinusdreef 2, 2300 RC Leiden, the Netherlands