Single-cell tracking of therapeutic cells using Laser Ablation–Inductively Coupled Plasma–Mass Spectrometry

Cellular therapy is emerging as a clinically viable strategy in the field of solid organ transplantation, where it is expected to reduce the dependency on conventional immunosuppression. This has produced a demand for highly sensitive methods to monitor the persistence and tissue distribution of administered cells in vivo. However, tracking cells presents significant challenges. In many cases transplanted cells are autologous with the immune system of the transplant recipient, and hence are invisible to typical methods of detection. To enable their differentiation, the cells must be labelled with a suitable, non-toxic and long lifetime label, prior to their administration to patients. In addition, administered cells represent only a small fraction of the recipient’s endogenous cells, which necessitates the use of an extremely sensitive detection method. Laser ablation – inductively coupled plasma – mass spectrometry (LA-ICP-MS) is an exquisitely sensitive analytical technique, capable of imaging trace elements in complex samples, at high spatial resolution. [Continues.

Doubling sensitivity in multicollector ICPMS using high-efficiency, rapid response Laser Ablation Technology

The introduction of rapid response laser ablation cells and sample transport technologies to laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) has enabled signal pulse durations for a single laser ablation shot of less than 10 ms. These developments have resulted in marked improvements in analytical throughput, resolution, and sensitivity vital for the generation of large, highly spatially resolved elemental maps. The focus on mapping, particularly bioimaging, has obscured the possibility of applying the sensitivity advantage of rapid response technologies to other LA-ICPMS applications, such as high-precision isotope ratio analysis on multicollector (MC) ICPMS. In this work a commercially available rapid response sample transport system and a conventional configuration were compared for LA-MC-ICPMS analysis. Ablation of known reference materials demonstrated “sensitivity” or sample ion yield of 7–9% using the rapid response sample transport system, more than double that for the conventional setup. This increase in efficiency was demonstrated to improve precision for the Pb isotope ratio analysis of the MPI-DING reference glasses and improve the spatial resolution of Hf isotope ratio analysis of reference zircon

Needles in haystacks: using fast-response LA chambers and ICP-TOF-MS to identify asbestos fibres in malignant mesothelioma models

Malignant mesothelioma is an aggressive cancer associated with exposure to asbestos. Diagnosis of mesothelioma and other related lung diseases remains elusive due to difficulties surrounding identification and quantification of asbestos fibres in lung tissue. This article presents a laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) method to identify asbestos fibres in cellular models of mesothelioma. Use of a high-speed laser ablation system enabled rapid imaging of the samples with a lateral resolution of 3 μm, whilst use of a prototype time-of-flight ICP-MS provided pseudo-simultaneous detection of the elements between mass 23 (Na) and mass 238 (U). Three forms of asbestos fibre (actinolite, amosite and crocidolite) were distinguished from a non-asbestos control (wollastonite) based on their elemental profile, which demonstrated that LA-ICP-MS could be a viable technique for identification of asbestos fibres in clinical research samples

Laser ablation inductively coupled plasma mass spectrometry as a novel clinical imaging tool to detect asbestos fibres in malignant mesothelioma.

RATIONALE: Malignant pleural mesothelioma is an extremely aggressive and incurable malignancy associated with prior exposure to asbestos fibres. Difficulties remain in relation to early diagnosis, notably due to impeded identification of asbestos in lung tissue. This study describes a novel laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) imaging approach to identify asbestos within mesothelioma models with clinical significance. METHODS: Human mesothelioma cells were exposed to different types of asbestos fibres and prepared on plastic slides for LA-ICP-MS analysis. No further sample preparation was required prior to analysis, which was performed using an NWR Image 266nm laser ablation system coupled to an Element XR sector-field ICP mass spectrometer, with a lateral resolution of 2 μm. Data was processed using LA-ICP-MS ImageTool v1.7 with the final graphic production made using DPlot Software. RESULTS: Four different mineral fibres were successfully identified within the mesothelioma samples based on some of the most abundant elements that make up these fibres (Si, Mg and Fe). Using LA-ICP-MS as an imaging tool provided information on the spatial distribution of the fibres at cellular level, which is essential in asbestos detection within tissue samples. Based on the metal counts generated by the different types of asbestos, different fibres can be identified based on shape, size, and elemental composition. Detection of Ca was attempted but requires further optimisation. CONCLUSION: Asbestos fibres detection in the lung tissues is very useful, if not necessary, to complete the pathological diagnosis of asbestos-related malignancies in medicolegal field. For the first time, this study demonstrates the successful application of LA-ICP-MS imaging to identify asbestos fibres and other mineral fibres within mesothelioma samples. Ultimately, high-resolution, fast-speed LA-ICP-MS analysis has the potential to be integrated into clinical workflow to aid earlier detection and stratification of mesothelioma patient samples

Single cell tracking of gadolinium labeled CD4(+) T cells by laser ablation inductively coupled plasma mass spectrometry

Cellular therapy is emerging as a promising alternative to conventional immunosuppression in the fields of haematopoietic stem cell (HSC) transplantation, autoimmune disease and solid organ transplantation. Determining the persistence of cell-based therapies in vivo is crucial to understanding their regulatory function and requires the combination of an extremely sensitive detection technique and a stable, long-lifetime cell labelling agent. This paper reports the first application of laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to perform single cell detection of T cell populations relevant to cellular immunotherapy. Purified human CD4+ T cells were labelled with commercially available Gd-based MRI contrast agents, Omniscan® and Dotarem®, which enabled passive loading of up to 108 Gd atoms per cell. In mixed preparations of labelled and unlabelled cells, LA-ICP-MS was capable of enumerating labelled cells at close to the predicted ratio. More importantly, LA-ICP-MS single cell analysis demonstrated that the cells retained sufficient label to remain detectable for up to 10 days post-labelling both in vitro and in vivo in an immunodeficient mouse model

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

High-Speed, Integrated Ablation Cell and Dual Concentric Injector Plasma Torch for Laser Ablation-Inductively Coupled Plasma Mass Spectrometry

In recent years, laser ablation-inductively coupled plasma mass spectrometry (LA-ICPMS) has gained increasing importance for biological analysis, where ultratrace imaging at micrometer resolution is required. However, while undoubtedly a valuable research tool, the washout times and sensitivity of current technology have restricted its routine and clinical application. Long periods between sampling points are required to maintain adequate spatial resolution. Additionally, temporal signal dispersion reduces the signal-to-noise ratio, which is a particular concern when analyzing discrete samples, such as individual particles or cells. This paper describes a novel, two-volume laser ablation cell and integrated ICP torch designed to minimize aerosol dispersion for fast, efficient sample transport. The holistic design utilizes a short, continuous diameter fused silica conduit, which extends from the point of ablation, through the ICP torch, and into the base of the plasma. This arrangement removes the requirement for a dispersive component for argon addition, and helps to keep the sample on axis with the ICP cone orifice. Hence, deposition of sample on the cones is theoretically reduced with a resulting improvement in the absolute sensitivity (counts per unit mole). The system described here achieved washouts of 1.5, 3.2, and 4.9 ms for NIST 612 glass, at full width half, 10%, and 1% maximum, respectively, with an 8–14-fold improvement in absolute sensitivity, compared to a single volume ablation cell. To illustrate the benefits of this performance, the system was applied to a contemporary bioanalytical challenge, specifically the analysis of individual biological cells, demonstrating similar improvements in performance

Fluorinated N‑Heterocyclic Carbene Silver(I) Complexes with High Cancer Cell Selectivity

This work presents the synthesis of five new functionalized (benz)imidazolium N-heterocyclic (NHC) ligands (L) and four new (benz)imidazole silver(I) NHC (Ag(I)-NHC) complexes of mononuclear [Ag(L) 2 ](PF 6 ) or binuclear [Ag 2 (L) 2 ](PF 6 ) 2 type. The complexes have been fully characterized, including single crystal X-ray diffraction of three new structures. The complexes and their corresponding free NHC ligands have been screened against breast cancer and noncancerous cell lines, showing the mononuclear benzimidazole complex has the highest activity, while the binuclear benzimidazole complex has the highest cancer cell selectivity. The silver uptake was measured by ICP-MS and highlights a strong link between cytotoxicity and cellular uptake. DNA interaction studies, molecular docking, and evaluation of reactive oxygen species (ROS) have been conducted for the most promising complexes to identify modes of action. Overall, the binuclear benzimidazole complex is the most selective and promising candidate against the MDA-MD-231 (breast cancer) cell line and has potential to be developed for treatment of late-stage breast cancers