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1.
Quantitative aspects of ToF-SIMS analysis of metals and alloys in a UHV, O2 and H2 atmosphere
Jernej Ekar, Saša Kos, Janez Kovač, 2024, original scientific article

Abstract: Although secondary ion mass spectrometry (SIMS) is a versatile method used in the fields of surface analysis, depth profiling and elemental and molecular mapping, it also lacks quantification capabilities. The main reason for this is the matrix effect, which influences the ionization yield of secondary ions with respect to the substrate from which the analyzed compounds originate. There are several approaches to reduce the matrix effect, and gas flooding is one of the easiest methods to apply. In this work, we have investigated the possibilities of the ToF-SIMS method for the quantification of selected metals and alloys containing these metals in different ratios by reducing the matrix effect in the presence of different atmospheres. The measurements were performed in the ultra-high vacuum (UHV) environment, H2 and O2 atmospheres. H2 flooding shows the most significant improvements compared to the UHV analysis, while O2 is also promising but has some limitations. Improvements are most evident for the transition metals Ti, Cr, Fe, Co and Ni employed in our study, while the p-block elements such as Al and Si do not change so extensively. The deviations from the true atomic ratios of selected transition metals in different alloys reach a maximum of only 46 % when analyzed in the H2 atmosphere. In contrast, these values are 66 and 228 % for the O2 atmosphere and UHV environment, respectively. Our results suggest that gas adsorption and consequent formation of a new matrix on the surface, especially in the case of hydrogen, reduces the differences between the different chemical environments and electronic structures of the surface. In this way, the quantitative aspects of the SIMS method can be improved.
Keywords: ToF-SIMS quantification, H2 and O2 gas flooding, matrix effect reduction, cluster secondary ions
Published in DiRROS: 15.05.2024; Views: 21; Downloads: 9
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2.
Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR
Alexandra Bogožalec Košir, Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory, David Dobnik, 2023, original scientific article

Abstract: The proliferation of genetically modified organisms (GMOs) presents challenges to GMO testing laboratories and policymakers. Traditional methods, like quantitative real-time PCR (qPCR), face limitations in quantifying the increasing number of GMOs in a single sample. Digital PCR (dPCR), specifically multiplexing, offers a solution by enabling simultaneous quantification of multiple GMO targets. This study explores the use of the Naica six-color Crystal dPCR platform for quantifying five GM soybean lines within a single six-plex assay. Two four-color assays were also developed for added flexibility. These assays demonstrated high specificity, sensitivity (limit of detection or LOD < 25 copies per reaction) and precision (bias to an estimated copy number concentration <15%). Additionally, two approaches for the optimization of data analysis were implemented. By applying a limit-of-blank (LOB) correction, the limit of quantification (LOQ) and LOD could be more precisely determined. Pooling of reactions additionally lowered the LOD, with a two- to eight-fold increase in sensitivity. Real-life samples from routine testing were used to confirm the assays’ applicability for quantifying GM soybean lines in complex samples. This study showcases the potential of the six-color Crystal dPCR platform to revolutionize GMO testing, facilitating comprehensive analysis of GMOs in complex samples.
Keywords: digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology
Published in DiRROS: 29.03.2024; Views: 144; Downloads: 60
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