ToF SIMS Research Articles

An insight into the origin of elemental chromium in the lacquer of Qin terracotta warriors

The origin of elemental chromium for the archaeological weapons from the pits of Qin terracotta warriors in China has been highly controversial. Although previous studies have highlighted that the chromium on the surface of weapon originated from the contamination of surrounding lacquer, the exact origin of chromium in the lacquer remains unclear. In this work, the measurement by inductively coupled plasma-mass spectrometer (ICP-MS) firstly confirmed that the elemental chromium was indeed contained in the archaeological Qin original lacquer. Nevertheless, the amount of elemental chromium in the Qin lacquer was as low as 0.0759 μg/mg, disclosing that it was impossible to artificially add extra refined chromium-containing substance to the lacquer in the preparation of the terracotta warriors. The soil from the archaeological site of Qin lacquer was found to have a chromium amount of 0.0660 μg/mg by ICP-MS. After the hygrothermal and soil-buried aging cycles for the lab-prepared lacquer, the surface and depth elemental analyses by time of flight-secondary ion mass spectrometer (TOF–SIMS) showed a gradient distribution of elemental chromium from the surface to interior of aged lacquer, indicating the migration and enrichment behavior of elemental chromium from the burial soil towards the lacquer. To explore the migration mechanism of elemental chromium, fluorescence imaging technique was employed in combination with Fourier transform infrared spectrometry (FT-IR) and X-ray photoelectron spectroscopy (XPS) characterizations. The results revealed that catechol-containing fragments were formed during hygrothermal and soil-buried aging of lacquer and consequently coordinated with chromium ions, inducing the migration of elemental chromium towards the lacquer.

Mechanism and experimental study of in situ metallization on the surface of silver ion conductive glass

AbstractIn situ metallization of Ag+ conductive glass was successfully achieved by applying electric and temperature fields to the glass to ensure the controlled migration of ions inside the glass. The transport and kinetic characteristics of Ag+ under the combined effect of electric field and temperature field were investigated by calculating and analyzing Ag+ diffusion coefficient. The correlations between ion transport and current were characterized with potential energy equations. The simulations and experiments of ion transport in the conducting glass were implemented, which revealed the mechanisms of ion transport and interfacial precipitation within the conducting glass. Further more, the existence of Ag+ deprivation layer was confirmed and it was clarified that the depth of the deprivation layer reached 30 µm under thermal polarization conditions of 60 V and 300°C. The microscopic morphology of in situ silver deposition layer was observed using SEM and TOF–SIMS. In addition, It was determined that the in situ metallization of conductive glass under the polarization condition of 60 V at 250°C was the best, and the thickness of deposited silver dendrites reached 1000 nm in the side of cathodically. This paper provides theoretical and experimental references for the optimization of the in situ metallization process for the ion conductive glass.

29‐4: TOF SIMS for OLED Film 3D Detection and Real‐Time Failure Analysis

By analyzing OLED film layers using the TOF SIMS method with 3D imaging capability, we can effectively improve the detection of OLED defects and understand the constitutive relationship. This inspection method can be integrated into the production line to realize process improvement and defect interception during mass production.

Microstructures, geochemistry, and geochronology of mica fish: Review and advances

Shear zones are one of the most conspicuous structures in orogenic and rifting settings, accommodating high strain, rock displacement and influencing magma emplacement and fluid flow. Since shear zones may be simultaneously or sequentially affected by multiple processes involving mineral reactions, variations in pressure-temperature conditions, fluid-rock interaction, and diffusion, determining the timing of such structures has been one of the major challenges for modern geochronology. Although low- (up to lower greenschist facies) and high-temperature shear zones (above amphibolite facies) are well-dated through low- and high-closure temperature minerals, medium-temperature shear zones developed within the critical temperature window of ∼450–550 °C, in which conventional chronometers such as 40Ar/39Ar and Rb–Sr applied to mica fish may or may not record the timing of deformation for multiple reasons (e.g., grain size, cooling rate, mineral composition, fluid activity, deformation, neo- and recrystallization). Here, we review the current knowledge on the evolution of mica fish and the effect of deformation on its chemical and isotopic systems. We evaluate the effect on the widely deployed in situ 40Ar/39Ar technique. Furthermore, we demonstrate the potential to assess mica fish evolution applying high-spatial resolution microstructural and chemical mapping techniques such as electron backscatter diffraction (EBSD), time-of-flight secondary ion mass spectrometry (ToF–SIMS) and in situ Rb–Sr via triple quadrupole inductively coupled plasma mass spectrometry (TQ-ICP-MS) to a case study of medium-temperature mylonites from the well-characterized Taxaquara shear zone, SE Brazil. We show that mica fish display complex microstructures with variable strain intensity, commonly with low strain inner cores and high strain edges and along kink planes. Strain shadows in mica fish are commonly characterized by low-strain fine-grained muscovite, suggesting recrystallization coeval with ductile deformation. Despite being intensely deformed, muscovite fish Rb–Sr retain the protolith age (c. 600 Ma), whereas recrystallized fine-grained muscovite yields the timing of deformation (c. 550–540 Ma). Synthetic shear bands cross-cutting coarse-grained muscovite fish induce muscovite recrystallization consistent with their distinct chemistry, with recrystallized muscovite characterized by higher Fe–Mg and lower Na suggesting fluid-assisted recrystallization under lower temperature compared to the muscovite fish host. We propose that these shear bands across mica fish play an important role by accommodating grain size reduction and subsequent deformation, leading to the formation of smaller individual mica fish. Grain size reduction, likely enhanced by dynamic precipitation (i.e., coeval crystal-plastic deformation and dissolution-precipitation creep), appears as the key recrystallization mechanism that allows low strain muscovite in strain shadows and shear bands to record the timing of deformation in medium-temperature shear zones, consistent with qualitative Sr diffusion modelling.

Influence of A-site Deficiency and Ni/Co Ratio in B-site on Electrochemical Performance of (La0.25Sr0,25Ca0.45)yTi0.95Ni0.05-xCoxO3- d Anode

La0.25Sr0.25Ca0.45TiO3 -d (LSCT) is a perovskite (ABO3) type mixed ionic-electronic conductive (MIEC) oxide and has been proposed as an electrode material for high temperature fuel cell [1]. This material owing high conductivity, robustness in hydrocarbon fuels and significant amount of attention has been paid to improve the electrochemical activity [1, 2]. Doping of B-site with some d-metal cation (Ni, Co, Mn, V, Mo) has been demonstrated to improve the catalytic activity. One of the advantages of the MIEC conducting scaffold based electrodes is that the catalyst phase on the electrode surface can be kept to a minimum, usually less than 5 wt%, which minimizes any risks of physical damage during redox cycling [2].In this work, Ni/Co ratio and deficiency of A-site, of (La0.25Sr0.25Ca0.45)xTi0.95Ni0.05-yCoyO3 -d were varied. Electrical as well as electrochemical performance and chemical composition of LSCTNC surface was monitored. The electrochemical measurements of symmetric cells during 100 h tests show that small stochiometric changes in A-site significantly influence the activity and initial degradation rate of the electrode. The chemical and structural changes of the material surface have a key role on the electrochemical performance of the electrode [3].The electrode materials were analysed using XRD, TOF SIMS and electrochemical methods. XRD and TOF SIMS results for studied electrode powders showed significant dependence of the lattice parameters and electrode surface composition on the perovskite elemental composition. The results from impedance spectroscopy (measured at temperatures from 973 to 1123 K in H2 environment, at OCV) demonstrate a significant influence of the A-site deficiency and B-site composition on the electrochemical properties of studied electrodes. Robert Price, Mark Cassidy, Jan G. Grolig, Gino Longo, Ueli Weissen, Andreas Mai, John T. S. Irvine, Advanced Energy Materials, 11, 1 (2021). Paul A. Connor, Xiangling Yue, Cristian D. Savaniu, Robert Price, Georgios Triantafyllou, Mark Cassidy, Gwilherm Kerherve, David J. Payne, Robert C. Maher, Lesley F. Cohen, Rumen I. Tomov, Bartek A. Glowacki, Ramachandran Vasant Kumar, John T. S. Irvine, Advanced Energy Materials, 8, 1 (2018). Ove Korjus, Priit Möller, Kuno Kooser, Tanel Käämbre, Olga Volobujeva, Jaak Nerut, Sander Kotkas, Enn Lust, Gunnar Nurk, Journal of Power Sources, 494, 1 (2021).

Evidence for Glass–glass Interfaces in a Columnar Cu–Zr Nanoglass

AbstractComprehensive analyses of the atomic structure using advanced analytical transmission electron microscopy‐based methods combined with atom probe tomography confirm the presence of distinct glass–glass interfaces in a columnar Cu‐Zr nanoglass synthesized by magnetron sputtering. These analyses provide first‐time in‐depth characterization of sputtered film nanoglasses and indicate that glass–glass interfaces indeed present an amorphous phase with reduced mass density as compared to the neighboring amorphous regions. Moreover, dedicated analyses of the diffusion kinetics by time‐of‐flight secondary ion mass spectroscopy (ToF SIMS) prove significantly enhanced diffusivity, suggesting fast transport along the low density glass–glass interfaces. The present results further indicate that sputter deposition is a feasible technique for reliable production of nanoglasses and that some of the concepts proposed for this new class of glassy materials are applicable.

The depression mechanism on pyrite in a low-alkaline system with combined depressants: Experiment, HSC, DFT and ToF–SIMS studies

Depression of pyrite in a low-alkaline system has sparked soaring interests for the multi-metal sulfide minerals flotation recently. This study investigates effects of combined depressants (Ca(ClO)<sub>2</sub> and CaO) on pyrite flotation with butyl xanthate (KBX). Micro-flotation experiments indicate that the addition of 200 mg/L combined depressants (a mass ratios of CaO to Ca(ClO)<sub>2</sub> of 2:3) and 1.0×10<sup>−3</sup> mol/L KBX at pH 9.5 can effectively depresses the flotation of pyrite, and a minimum pyrite recovery rate of 12.5% is obtained. Basic thermodynamic evaluation results confirm the participation of Ca(ClO)2 significantly decrease the negative Gibbs free energies of pyrite oxidation reaction. Besides, the calcium species (Ca(OH)<sub>2</sub>, Ca<sup>2+</sup> and Ca(Cl)<sub>2</sub>) will spontaneously transform into CaCO<sub>3</sub>, and it is the ultimate dominant calcium species in the CO<sub>3</sub><sup>2-</sup> system. Density functional theory (DFT) results indicate that CaCO<sub>3</sub> can chemically adsorb onto the pyrite surface with an adsorption energy of –671.13 kJ/mol. The O1 and Ca atoms mainly contribute to the bonding process and are responsible for the stable adsorption of CaCO<sub>3</sub>. ToF-SIMS results provide strong evidence that the combined depressants increase the amount of hydrophilic species and decrease dixanthogen adsorption onto the pyrite surface. The thickness of the whole formed hydrophilic species is approximately 50 nm. Semiquantitative amounts of hydrophilic species follow the order of hydroxy calcium>iron carbonyl>calcium carbonate. Overall, hydrophilic species repulse adsorption of dixanthogen and significantly reduce the flotation performance of pyrite.

The spatial distribution of soluble organic matter and their relationship to minerals in the asteroid (162173) Ryugu

We performed in-situ analysis on a ~ 1 mm-sized grain A0080 returned by the Hayabusa2 spacecraft from near-Earth asteroid (162173) Ryugu to investigate the relationship of soluble organic matter (SOM) to minerals. Desorption electrospray ionization-high resolution mass spectrometry (DESI-HRMS) imaging mapped more than 200 CHN, CHO, CHO–Na (sodium adducted), and CHNO soluble organic compounds. A heterogeneous spatial distribution was observed for different compound classes of SOM as well as among alkylated homologues on the sample surface. The A0080 sample showed mineralogy more like an Ivuna-type (CI) carbonaceous chondrite than other meteorites. It contained two different lithologies, which are either rich (lithology 1) or poor (lithology 2) in magnetite, pyrrhotite, and dolomite. CHN compounds were more concentrated in lithology 1 than in lithology 2; on the other hand, CHO, CHO–Na, and CHNO compounds were distributed in both lithologies. Such different spatial distribution of SOM is likely the result of interaction of the SOM with minerals, during precipitation of the SOM via fluid activity, or could be due to difference in transportation efficiencies of SOMs in aqueous fluid. Organic-related ions measured by time-of-flight secondary ion mass spectrometry (ToF–SIMS) did not coincide with the spatial distribution revealed by DESI-HRMS imaging. This result may be because the different ionization mechanism between DESI and SIMS, or indicate that the ToF–SIMS data would be mainly derived from methanol-insoluble organic matter in A0080. In the Orgueil meteorite, such relationship between altered minerals and SOM distributions was not observed by DESI-HRMS analysis and field-emission scanning electron microscopy, which would result from differences of SOM formation processes and sequent alteration process on the parent bodies or even on the Earth. Alkylated homologues of CHN compounds were identified in A0080 by DESI-HRMS imaging as observed in the Murchison meteorite, but not from the Orgueil meteorite. These compounds with a large C number were enriched in Murchison fragments with abundant carbonate grains. In contrast, such relationship was not observed in A0080, implying different formation or growth mechanisms for the alkylated CHN compounds by interaction with fluid and minerals on the Murchison parent body and asteroid Ryugu.Graphical

TOF–SIMS analysis of mouse fibroblasts cells

In this paper, we have shown the results of the analysis by the SIMS method of mouse fibroblast cells (3T3-F44A2 line) grown on the silicon surface and fixed by using different chemicals. It was concluded that the standard method of dehydration and fixation of cells with the help of glutaraldehyde allows us to prepare relevant samples for analysis by the SIMS in ultrahigh vacuum. Ion images of chemical elements inside cells with a sufficiently high lateral resolution were obtained, allowing us to study, among other things, the internal structure of cells. The use of SIMS looks preferable for the analysis of cellular metabolism, cell differentiation, ischemia and other processes in cells and tissues grown using special medications and (or) reagents with an artificial isotopic composition of stable isotopes.

Evaluation of Short-Term Oxidation Mechanism of Laser Cladded Ni–Cr–C Coating on Titanium Aluminide Substrate

In the present investigation, a NiCr–Cr3C2 powder mixture was deposited on titanium aluminide substrate by laser cladding. The kinetics and mechanism of oxidation of the substrate and coating were studied at 800–1100 °C using XRD, SEM, ToF–SIMS and XPS characterization techniques. The results demonstrated that the Ni–Cr–C coating had improved oxidation resistance, as compared to the substrate. The oxidized surface of the coating was covered with Cr2O3 and NiCr2O4 phases at the early stages of the oxidation. The formation of NiCr2O4 could be attributed to the reaction of NiO (an oxidation product of the NiCr phase) and Cr2O3 (an oxidation product of the chromium carbide phase) during oxidation. Increasing the oxidation time at 900 °C led to the partial formation of CrO3 as a result of the oxidation of Cr2O3. It was also found that the parabolic rate constant ratio of $$\left( {K_{{\text{p}}}^{{{\text{substrate}}}} /K_{{\text{p}}}^{{{\text{coating}}}} } \right)$$ increased with the temperature, which showed, comparatively, the better oxidation resistance of the coating at higher temperatures.

Label-free sub-micrometer 3D imaging of ciprofloxacin in native-state biofilms with cryo-time-of-flight secondary ion mass spectrometry

High spatial resolution mass spectrometry imaging has been identified as a key technology needed to improve understanding of the chemical components that influence antibiotic resistance within biofilms, which are communities of micro-organisms that grow attached to a surface. Time-of-flight secondary ion mass spectrometry (ToF–SIMS) offers the unique ability for label-free 3D imaging of organic molecules with sub-micrometer spatial resolution and high sensitivity. Several studies of biofilms have been done with the help of ToF–SIMS, but none of those studies have shown 3D imaging of antibiotics in native-state hydrated biofilms with cell-level resolution. Because ToF–SIMS measurements must be performed in a high-vacuum environment, cryogenic preparation and analysis are necessary to preserve the native biofilm structure and antibiotic spatial distribution during ToF–SIMS measurements. In this study, we have investigated the penetration of the antibiotic ciprofloxacin into Bacillus subtilis biofilms using sub-micrometer resolution 3D imaging cryo-ToF–SIMS. B. subtilis biofilms were exposed to physiologically relevant levels of ciprofloxacin. The treated biofilms were then plunge-frozen in liquid propane and analyzed with ToF–SIMS under cryogenic conditions. Multivariate analysis techniques, including multivariate curve resolution (MCR) and inverse maximum signal factor (iMSF) denoising, were used to aid analysis of the data and facilitate high spatial resolution 3D imaging of the biofilm, providing individually resolved cells and spatially resolved ciprofloxacin intensity at “real world” concentrations.Graphical

Plasma-enabled superhydrophobic coatings on mild steel

This work demonstrates a new pathway to the direct on-surface fabrication of a superhydrophobic surface coating on mild steel. The coating was formed using dielectric barrier discharge (DBD) plasma to convert a liquid small-molecule precursor (1,2,4-tricholorobenzene) to a solid film via plasma-assisted on-surface polymerization. Plasma treatments were performed under a nitrogen atmosphere with a variety of power levels and durations. Samples were analysed by optical and scanning electron microscopy (SEM), energy-dispersive x-ray spectroscopy (EDS), time-of-flight secondary ion mass spectrometry (ToF–SIMS), Raman spectroscopy, optical profilometry, contact angle measurement, and potentiodynamic polarisation tests. Wettability of the films varied with the plasma parameters, and through the inclusion of graphene nanoplatelets in the precursor. High-dose plasma exposures of the nanoplatelet-containing precursor created superhydrophobic films with water contact angles above 150°. Potentiodynamic polarisation tests revealed that the superhydrophobic coating provided little or no corrosion protection.

Lithium isotope tracing in silicon-based electrodes using solid-state MAS NMR: a powerful comprehensive tool for the characterization of lithium batteries.

The introduction of lithiated components with different 7Li/6Li isotopic ratios, also called isotopic tracing, can give access to better understanding of lithium transport and lithiation processes in lithium-ion batteries. In this work, we propose a simple methodology based on high-resolution solid-state NMR for the determination of the 7Li/6Li ratio in silicon electrodes following different strategies of isotopic tracing. The 6Li and 7Li MAS NMR experiments allow obtaining resolved spectra whose spectral components can be assigned to different moieties of the materials. In order to measure the ratio of the 6Li/7Li NMR integrals, a silicon electrode with a natural 7Li/6Li isotope abundance was used as a reference. This calibration can then be used to determine the 7Li/6Li ratio of any similar samples. This method was applied to study the phenomena occurring at the interface between a silicon electrode and a labeled electrolyte, which is an essential step for isotopic tracing experiments in systems after the formation of the solid electrolyte interphase (SEI). Beyond the isotopic exchanges between the SEI and the electrolyte already observed in the literature, our results show that isotopic exchanges also involve Li-Si alloys in the electrode bulk. Within a 52-hour contact, the electrolyte labeling disappeared: isotopic concentrations of the electrolyte and electrode become practically homogenized. However, at the electrode level, different silicides are characterized by rather different isotopic enrichment. In the present work, ToF SIMS and liquid state NMR were also used to cross-check and discuss the solid-state NMR method we have proposed.

Evaluation of the relative potential for contact and doffing transmission of SARS-CoV-2 by a range of personal protective equipment materials

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)—the causative agent of coronavirus disease 2019 (COVID-19)—has caused a global public health emergency. Personal protective equipment (PPE) is the primary defence against viral exposure in healthcare and community settings. However, the surfaces of PPE materials may trap virus for contact transmission or through laden aerosols generated during removal of PPE, through cleaning or during movement. In this study, the relative efficacy of current PPE materials in terms of virion adsorption to materials and their antiviral potency, has been evaluated on a wide range of PPE for the first time, including four polymer glove types, two types of scrubs, apron material, a mask, visor and a selection of other commercial polymers and products. Although differences in virion adsorption to the test materials were observed, none of the existing polymer-based PPE resulted in more than tenfold reduction in the SARS-CoV-2 titre within either 10 min or 30 min contact period. The wettability and surface chemistry of the test materials were analysed to investigate any correlations with their surface physicochemical properties. While no correlation was found between wettability and viral retention under air flow challenge, one secondary ion of m/z 101.03 (+) and three secondary ions of m/z 31.98 (−), 196.93 (−) and 394.33 (+) in ToF–SIMS data of the test materials showed positive and negative correlations with the viral retention, respectively, which was identified by PLS regression model, suggesting that the surface chemistry plays a role in determining the extent of virion adsorption. Our findings outline the material aspects that influence the efficacy of current PPE against SARS-CoV-2 transmission and give suggestions on the development of novel simple polymer-based PPE for better infection protection.

Effect of Ammonium Sulfide on Sulfidization Flotation of Malachite

Recently, several studies have shown the positive effect of sulfidization flotation on malachite surfaces and its enhancing methods. Therefore, this paper was focused on the effect of ammonium sulfide and sodium sulfide on the sulfidization of malachite, respectively; this was investigated using different devices such as the micro-flotation tests, Zeta potential measurements, ToF–SIMS, XPS analysis, and FTIR. Thus, Fourier transform infrared spectroscopy results demonstrated that a new characteristic peak of Cu-S bonds was formed and adsorbed on malachite surfaces at 1694 cm−1, as confirmed by XPS analysis. Notably, malachite with ammonium sulfide ions had a significantly higher flotation recovery than malachite with an excess of sodium sulfide ions, as concerns of sulfidization types. Conclusively, all the experiments in this study confirmed that additional copper sulfide products were formed on the malachite surface, increasing the hydrophobicity of the malachite.

3‐2: Analyzing the Degradation Process of Quantum‐Dot LEDs (QLEDs) by Mass Spectometry

By utilizing time of flight secondary ion mass spectrometry (TOF SIMS), the degradation processes of quantum dot core components and the surface ligands were studied. Intentionally degraded QLED samples were prepared, and the TOF SIMS mass composition signals of each layer within the QLEDs are estimated. The result of the analysis gives possible structural information of the degraded species induced in the device, providing useful insights for further developments of highly efficient R/G/B quantum dot materials.

Silicon Contamination During Alloy Oxidation in Water Vapour at 650 °C

A model Si-free alloy, Fe-30Cr (wt.%), was exposed to Ar-10H2O mixture (in vol.%) at 650 °C, forming a protective Cr2O3 scale, which contained silica. Liquid water used to form water vapour in a bubbler was either deionised or distilled. In both cases, low-level water impurities of Si, F, Cl, S, and Ca matched those detected in the Cr2O3 scales by TEM/EDX and ToF–SIMS. Consideration of water droplet and residual solute particle sizes, together with Stokes’ Law shows that entrainment of Si in the gas stream is likely. Calculation of mass transfer rates from the gas to the growing chromia scale show that the amount of silica in the scale is accounted for. Many laboratory experiments could be affected in this way.

Understanding the effect of grinding media on the adsorption mechanism of cyanide to chalcopyrite surface by ToF–SIMS, XPS, contact angle, zeta potential and flotation

As the main copper-bearing sulfide minerals in cyanide tailings, how to improve the flotation recovery of chalcopyrite is of great significance to the comprehensive management of bulk solid waste. To obtain higher leaching rate of precious metals, fine grinding is required. However, the production of numerous interfering ions, accompanied by severe mechanical entrainment, leads to a series of problems in the grinding process. In this context, the correlation between surface adsorption, the occurrence of species on the chalcopyrite surface and the differences in grinding media in cyanidation system would be a key issue for process optimization. Scanning electron microscope–energy dispersive spectrometer (SEM–EDS) and contact angle measurements were performed on the chalcopyrite surface. The results showed that the use of ceramic media avoided excessive surface oxidation caused by the galvanic couple actions between the iron media and chalcopyrite. Zeta potential values revealed that the cyanide adsorbed on the chalcopyrite surface and reacted with positively charged substances, inhibiting the adsorption of collector, which was confirmed by microflotation. Surface chemistry analysis using X–ray photoelectron spectroscopy (XPS) and time–of–flight secondary ion mass spectrometry (ToF–SIMS) confirmed that the chemical environment of the chalcopyrite surface had been changed using the iron media, which accelerated the formation of FeOOH and enhanced the adsorption of CN– on the chalcopyrite surface. It was further demonstrated by ToF-SIMS that the ceramic media exhibited the excellent grinding performance, which excluded the insignificant contribution of impurity elements (mainly Fe). This study provides important surface chemical evidence to gain an in-depth understanding of the effect of grinding environment on the mechanism of cyanide adsorption on the chalcopyrite surface, which plays a crucial role in promoting the possibility of the comprehensive utilization of the cyanide tailings.

Subcellular detection of PEBCA particles in macrophages: combining darkfield microscopy, confocal Raman microscopy, and ToF–SIMS analysis

The detection of biomedical organic nanocarriers in cells and tissues is still an experimental challenge. Here we developed an imaging strategy for the label-free detection of poly (ethylbutyl cyanoacrylate) (PEBCA) particles. Experiments were carried out with phagocytic NR8383 macrophages exposed to non-toxic and non-activating concentrations of fluorescent (PEBCA NR668 and PEBCA NR668/IR), non-fluorescent (PEBCA), and cabazitaxel-loaded PEBCA particles (PEBCA CBZ). Exposure to PEBCA NR668 revealed an inhomogeneous particle uptake similar to what was obtained with the free modified Nile Red dye (NR668). In order to successfully identify the PEBCA-loaded cells under label-free conditions, we developed an imaging strategy based on enhanced darkfield microscopy (DFM), followed by confocal Raman microscopy (CRM) and time-of-flight secondary ion mass spectrometry (ToF–SIMS). Nitrile groups of the PEBCA matrix and PEBCA ions were used as suitable analytes for CRM and ToF–SIMS, respectively. Masses found with ToF–SIMS were further confirmed by Orbitrap-SIMS. The combined approach allowed to image small (< 1 µm) PEBCA-containing phagolysosomes, which were identified as PEBCA-containing compartments in NR8383 cells by electron microscopy. The combination of DFM, CRM, and ToF–SIMS is a promising strategy for the label-free detection of PEBCA particles.Graphical

Lipofuscin Granule Bisretinoid Oxidation in the Human Retinal Pigment Epithelium forms Cytotoxic Carbonyls.

Age-related macular degeneration (AMD) is the primary cause of central blindness among the elderly. AMD is associated with progressive accumulation of lipofuscin granules in retinal pigment epithelium (RPE) cells. Lipofuscin contains bisretinoid fluorophores, which are photosensitizers and are phototoxic to RPE and neuroretinal cells. In the presence of oxygen, bisretinoids are also oxidized, forming various products, consisting primarily of aldehydes and ketones, which are also potentially cytotoxic. In a prior study, we identified that in AMD, bisretinoid oxidation products are increased in RPE lipofuscin granules. The purpose of the present study was to determine if these products were toxic to cellular structures. The physicochemical characteristics of bisretinoid oxidation products in lipofuscin, which were obtained from healthy donor eyes, were studied. Raman spectroscopy and time-of-flight secondary ion mass spectrometry (ToF–SIMS) analysis identified the presence of free-state aldehydes and ketones within the lipofuscin granules. Together, fluorescence spectroscopy, high-performance liquid chromatography, and mass spectrometry revealed that bisretinoid oxidation products have both hydrophilic and amphiphilic properties, allowing their diffusion through lipofuscin granule membrane into the RPE cell cytoplasm. These products contain cytotoxic carbonyls, which can modify cellular proteins and lipids. Therefore, bisretinoid oxidation products are a likely aggravating factor in the pathogenesis of AMD.