RESUMEN
The ionization edges encoded in the electron energy loss spectroscopy (EELS) spectra enable advanced material analysis including composition analyses and elemental quantifications. The development of the parallel EELS instrument and fast, sensitive detectors have greatly improved the acquisition speed of EELS spectra. However, the traditional way of core-loss edge recognition is experience based and human labor dependent, which limits the processing speed. So far, the low signal-noise ratio and the low jump ratio of the core-loss edges on the raw EELS spectra have been challenging for the automation of edge recognition. In this work, a convolutional-bidirectional long short-term memory neural network (CNN-BiLSTM) is proposed to automate the detection and elemental identification of core-loss edges from raw spectra. An EELS spectral database is synthesized by using our forward model to assist in the training and validation of the neural network. To make the synthesized spectra resemble the real spectra, we collected a large library of experimentally acquired EELS core edges. In synthesize the training library, the edges are modeled by fitting the multi-Gaussian model to the real edges from experiments, and the noise and instrumental imperfectness are simulated and added. The well-trained CNN-BiLSTM network is tested against both the simulated spectra and real spectra collected from experiments. The high accuracy of the network, 94.9%, proves that, without complicated preprocessing of the raw spectra, the proposed CNN-BiLSTM network achieves the automation of core-loss edge recognition for EELS spectra with high accuracy.
Asunto(s)
Aprendizaje Profundo , Humanos , Redes Neurales de la Computación , Espectroscopía de Pérdida de Energía de Electrones , Automatización , Memoria a Largo PlazoRESUMEN
We devise a new kind of experiment that extends the technology of electron energy loss spectroscopy to probe (supra-)molecular systems: by using an electron beam in a configuration that avoids molecular damage and a very recently introduced electron optics setup for the analysis of the outcoming electrons, one can obtain information on the spatial features of the investigated excitations. Physical insight into the proposed experiment is provided by means of a simple but rigorous model to obtain the transition rate and selection rule. Numerical simulations of DNA G-quadruplexes and other biomolecular systems, based on time dependent density functional theory calculations, point out that the conceived new technique can probe the multipolar components and even the chirality of molecular transitions, superseding the usual optical spectroscopies for those cases that are problematic, such as dipole-forbidden transitions, at a very high spatial resolution.
Asunto(s)
ADN/química , Teoría Funcional de la Densidad , Electrones , Transporte de Electrón , G-Cuádruplex , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
Electron energy-loss spectroscopy was used to characterize the boron profile and its coordination (BIII and BIV), along the complex alteration layer of glass samples altered for 511 days at 50 °C in solution containing FeCl2, MgCl2 and/or CaCl2. To reach this goal, the impact of both TEM operating conditions and sample preparation on the determination of the boron coordination was first studied using mineralogical and pristine glasses reference samples. Then, the boron concentration profiles were characterized in the glass alteration layer. These profiles were found to be S-shaped with a thickness around forty nanometers. The proportion of BIII was found to decrease with the boron total concentration (from the pristine glass to the gel layer), which suggests a higher bonding strength for BIV bonds than that of BIII bonds under the alteration conditions. These findings are of tremendous interest to advance further in the understanding of glass alteration mechanisms.
Asunto(s)
Boro/química , Vidrio/química , Espectroscopía de Pérdida de Energía de Electrones/métodos , Vidrio/análisis , Ensayo de Materiales , Minerales/análisisRESUMEN
Objective: The aim of the study was to fabricate and morphologically characterize ultrafine Polyetherimide fibers (PEI) associated with Polymethylmethacrylate (PMMA) PP (group formed by the association of PEI with PMMA), produced by the electrospinning process. Material and Methods: A solution of PEI (0.562 g) + PMMA (0.377 g) dissolved in 2.5 mL of chloroform, 0.85 mL of Dimethylformamide (DMF) and 0.85 mL of 1.1.2.2 Tetrachloroethane (TCE) was prepared. For the electrospinning process, different continuous voltages (10 to 18 kV) and two different distances (8 and 12 cm) between the needle tip and the collecting apparatus were used, giving rise to 6 distinct groups of ultrafine fibers (PP 1 to 6) that were observed in Scanning Electron Microscopy to check for defects and calculate the average diameter of the fibers. Results: The best parameter, the parameter that was most effective for the production of fibers, observed was subjected to Energy Dispersion X-ray Spectroscopy (EDS), X-ray Diffraction (XRD) and Contact Angle Analysis tests. The data were analyzed using the ANOVA and Tukey test (p <0.05). From the comparative analysis of the pre-established parameters, the pattern of PP4 ultrafine fibers was shown to be more effective. Conclusion: The PP4 standard (13 kV 12 cm) had an average diameter of 0.37 µm. An adequate parameter to electrospinning was able to produce ultrafine fibers of PMMA/PEI (AU)
Introdução: O objetivo do estudo foi sintetizar e caracterizar morfologicamente fibras ultrafinas de Polieterimida (PEI) associadas ao Polimetilmetacrilato (PMMA) - PP (grupo formado pela associação de PEI com PMMA), produzidas pelo processo de eletrofiação. Material e Métodos: Foi preparada uma solução de PEI (0,562 g) + PMMA (0,377 g) dissolvido em 2,5 mL de clorofórmio, 0,85 mL de Dimetilformamida (DMF) e 0,85 mL de 1.1.2.2 Tetracloroetano (TCE). Para o processo de eletrofiação, foram utilizadas diferentes tensões contínuas (10 a 18 kV) e duas distâncias diferentes (8 e 12 cm) entre a ponta da agulha e o aparelho coletor, dando origem a 6 grupos distintos de fibras ultrafinas (PP 1 a 6) que foram observados em Microscopia Eletrônica de Varredura para verificar defeitos e calcular o diâmetro médio das fibras. Resultados: O melhor parâmetro, o parâmetro mais eficaz para a produção de fibras, observado foi submetido aos testes de Espectroscopia de Dispersão de Energia (EDS), Difração de Raios X (DRX) e Análise do Ângulo de Contato. Os dados foram analisados pela ANOVA e teste de Tukey (p <0,05). A partir da análise comparativa dos parâmetros pré-estabelecidos, o padrão das fibras ultrafinas PP4 mostrou-se mais eficaz. Conclusão: O padrão PP4 (13 kV - 12 cm) apresentou diâmetro médio de 0,37 µm. Um parâmetro adequado para eletrofiação foi capaz de produzir fibras ultrafinas de PMMA / PEI. (AU)
Asunto(s)
Microscopía Electrónica de Rastreo , Polimetil Metacrilato , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
Idiopathic kidney stones originate mainly from calcium phosphate deposits at the tip of renal papillae, known as Randall's plaques (RPs), also detected in most human kidneys without stones. However, little is known about the mechanisms involved in RP formation. The localization and characterization of such nanosized objects in the kidney remain a real challenge, making their study arduous. This study provides a nanoscale analysis of the chemical composition and morphology of incipient RPs, characterizing in particular the interface between the mineral and the surrounding organic compounds. Relying on data gathered from a calculi collection, the morphology and chemical composition of incipient calcifications in renal tissue were determined using spatially resolved electron energy-loss spectroscopy. We detected microcalcifications and individual nanocalcifications found at some distance from the larger ones. Strikingly, concerning the smaller ones, we show that two types of nanocalcifications coexist: calcified organic vesicles and nanometric mineral granules mainly composed of calcium phosphate with carbonate in their core. Interestingly, some of these nanocalcifications present similarities with those reported in physiological bone or pathological cardiovascular biominerals, suggesting possible common formation mechanisms. However, the high diversity of these nanocalcifications suggests that several mechanisms may be involved (nucleation on a carbonate core or on organic compounds). In addition, incipient RPs also appear to present specific features at larger scales, revealing secondary calcified structures embedded in a fibrillar organic material. Our study proves that analogies exist between physiological and pathological biominerals and provides information to understand the physicochemical processes involved in pathological calcification formation.
Asunto(s)
Fosfatos de Calcio/análisis , Electrones , Cálculos Renales/diagnóstico por imagen , Médula Renal/diagnóstico por imagen , Nanotecnología , Espectroscopía de Pérdida de Energía de Electrones , Fosfatos de Calcio/metabolismo , Humanos , Cálculos Renales/química , Cálculos Renales/metabolismo , Médula Renal/química , Médula Renal/metabolismo , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Transmission electron microscopic (TEM) images of ion-milled bovid cortical bone cut approximately normal to the axes of fibrils show that mineral occurs in the form of plates surrounding and laying between circular or elliptical features about 50â¯nm in diameter. The classification of these features as either pores or collagen fibrils is highly debated. Electron energy loss spectroscopy (EELS) mapping of these features in ion milled sections shows that they are lacking significant amounts of mineral or collagen, although their appearance suggests that they are cross sections of collagen fibrils. However, analogous sections prepared using an ultramicrotome show that, while these circular features show reduced concentrations of calcium and phosphorus, some of them contain quantities of carbon and nitrogen in bonding states comparable to the composition of collagen. This work demonstrates that the observed circular features are sections of collagen fibrils, but that bombardment by argon ions during broad beam ion milling destroys the collagen and associated gap-zone mineral.
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Colágeno/química , Hueso Cortical/ultraestructura , Minerales/análisis , Espectroscopía de Pérdida de Energía de Electrones , Animales , Bovinos , Hueso Cortical/química , Tomografía con Microscopio Electrónico , Femenino , Manejo de EspecímenesRESUMEN
"Biased" G protein-coupled receptor (GPCR) agonists preferentially activate pathways mediated by G proteins or ß-arrestins. Here, we use double electron-electron resonance spectroscopy to probe the changes that ligands induce in the conformational distribution of the angiotensin II type I receptor. Monitoring distances between 10 pairs of nitroxide labels distributed across the intracellular regions enabled mapping of four underlying sets of conformations. Ligands from different functional classes have distinct, characteristic effects on the conformational heterogeneity of the receptor. Compared to angiotensin II, the endogenous agonist, agonists with enhanced Gq coupling more strongly stabilize an "open" conformation with an accessible transducer-binding site. ß-arrestin-biased agonists deficient in Gq coupling do not stabilize this open conformation but instead favor two more occluded conformations. These data suggest a structural mechanism for biased ligand action at the angiotensin receptor that can be exploited to rationally design GPCR-targeting drugs with greater specificity of action.
Asunto(s)
Angiotensinas/metabolismo , Receptor de Angiotensina Tipo 1/metabolismo , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Antagonistas de Receptores de Angiotensina/metabolismo , Arrestinas/metabolismo , Línea Celular , Humanos , Ligandos , Conformación Proteica , Receptores de Angiotensina/metabolismo , Receptores Acoplados a Proteínas G/agonistas , Receptores Acoplados a Proteínas G/metabolismo , Transducción de Señal , Espectroscopía de Pérdida de Energía de Electrones/métodos , beta-Arrestinas/metabolismoRESUMEN
Boron has been employed in materials science as a marker for imaging specific structures by electron energy loss spectroscopy (EELS) or secondary ion mass spectrometry (SIMS). It has a strong potential in biological analyses as well; however, the specific coupling of a sufficient number of boron atoms to a biological structure has proven challenging. Herein, we synthesize tags containing closo-1,2-dicarbadodecaborane, coupled to soluble peptides, which were integrated in specific proteins by click chemistry in mammalian cells and were also coupled to nanobodies for use in immunocytochemistry experiments. The tags were fully functional in biological samples, as demonstrated by nanoSIMS imaging of cell cultures. The boron signal revealed the protein of interest, while other SIMS channels were used for imaging different positive ions, such as the cellular metal ions. This allows, for the first time, the simultaneous imaging of such ions with a protein of interest and will enable new biological applications in the SIMS field.
Asunto(s)
Compuestos de Boro/síntesis química , Sondas Moleculares/síntesis química , Nanopartículas/química , Péptidos/química , Proteínas/análisis , Compuestos de Boro/metabolismo , Línea Celular , Química Clic , Imagen Molecular/métodos , Sondas Moleculares/metabolismo , Proteínas/inmunología , Espectrometría de Masa de Ion Secundario , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
Understanding the biomineralization pathways in living biological species is a grand challenge owing to the difficulties in monitoring the mineralization process at sub-nanometer scales. Here, we monitored the nucleation and growth of magnetosome nanoparticles in bacteria and in real time using a transmission electron microscope (TEM). To enable biomineralization within the bacteria, we subcultured magnetotactic bacteria grown in iron-depleted medium and then mixed them with iron-rich medium within graphene liquid cells (GLCs) right before imaging the bacteria under the microscope. Using in situ electron energy loss spectroscopy (EELS), the oxidation state of iron in the biomineralized magnetosome was analysed to be magnetite with trace amount of hematite. The increase of mass density of biomineralized magnetosomes as a function of incubation time indicated that the bacteria maintained their functionality during the in situ TEM imaging. Our results underpin that GLCs enables a new platform to observe biomineralization events in living biological species at unprecedented spatial resolution. Understanding the biomineralization processes in living organisms facilitates the design of biomimetic materials, and will enable a paradigm shift in understanding the evolution of biological species.
Asunto(s)
Biomineralización/fisiología , Grafito/química , Magnetosomas/química , Magnetosomas/metabolismo , Magnetospirillum/metabolismo , Microscopía Electrónica de Transmisión , Nanopartículas/química , Medios de Cultivo/química , Compuestos Férricos/metabolismo , Óxido Ferrosoférrico/metabolismo , Hierro/química , Hierro/metabolismo , Magnetosomas/ultraestructura , Magnetospirillum/ultraestructura , Nanopartículas/metabolismo , Nanopartículas/ultraestructura , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
In this chapter, we highlight the applications of electron microscopes (EMs) in nanotoxicity assessment. EMs can provide detailed information about the size and morphology of nanomaterials (NMs), their localization in cells and tissues, the nano-bio interactions, as well as the ultrastructural changes induced by NMs exposure. Here, we share with the readers how we prepare the tissue sample, and the different types of EMs used among the nanotoxicologists. It is possible to deploy conventional EMs along or in combination with other analytical techniques, such as electron energy loss spectroscopy (EELS), energy dispersive X-ray spectroscopy (EDS or EDX), and TEM-assisted scanning transmission X-ray microscopy (STXM), toward further elemental and chemical characterization. Appropriate images are inserted to illustrate throughout this chapter.
Asunto(s)
Técnicas de Preparación Histocitológica/métodos , Microscopía Electrónica de Transmisión de Rastreo/métodos , Nanopartículas/toxicidad , Espectrometría por Rayos X/métodos , Espectroscopía de Pérdida de Energía de Electrones/métodos , Animales , Línea Celular , Técnicas de Preparación Histocitológica/instrumentación , Humanos , Ratones , Microscopía Electrónica de Transmisión de Rastreo/instrumentación , Espectrometría por Rayos X/instrumentación , Espectroscopía de Pérdida de Energía de Electrones/instrumentaciónRESUMEN
The binding site of the extrinsic protein PsbP in plant photosystem II was mapped by pulsed electron-electron double resonance, using mutant spinach PsbP (Pro20Cys, Ser82Cys, Ala111Cys, and Ala186Cys) labeled with 4-maleimido-TEMPO (MSL) spin label. The distances between the spin label and the Tyr160 neutral radical (YD) in PsbD, the D2 subunit of plant photosystem II, were 50.8⯱â¯3.5â¯Å, 54.9⯱â¯4.0â¯Å, 57.8⯱â¯4.9â¯Å, and 58.4⯱â¯14.1â¯Å, respectively. The geometry inferred from these distances was fitted to the PsbP crystal structure (PDB: 4RTI) to obtain the coordinates of YD relative to PsbP. These coordinates were then fitted under boundary conditions to the structure of cyanobacterial photosystem II (PDB: 4UB6), by rotating on Euler angles centered at fixed YD coordinates. The result proposed two models which show possible acidic amino acid residues in CP43, CP47 and D2 that can bind the basic amino acids Arg48, Lys143, and Lys160 in PsbP.
Asunto(s)
Complejo de Proteína del Fotosistema II/química , Spinacia oleracea/enzimología , Sustitución de Aminoácidos , Mutación Missense , Complejo de Proteína del Fotosistema II/genética , Complejo de Proteína del Fotosistema II/metabolismo , Espectroscopía de Pérdida de Energía de Electrones , Spinacia oleracea/genéticaRESUMEN
Iron (Fe) is an essential metal involved in a wide spectrum of physiological functions. Sub-cellular characterization of the size, composition, and distribution of ferritin(iron) can provide valuable information on iron storage and transport in health and disease. In this study we employ magnetic force microscopy (MFM), transmission electron microscopy (TEM), and electron energy loss spectroscopy (EELS) to characterize differences in ferritin(iron) distribution and composition across injured and non-injured tissues by employing a rodent model of spinal cord injury (SCI). Our biophysical and ultrastructural analyses provide novel insights into iron distribution which are not obtained by routine biochemical stains. In particular, ferritin(iron) rich lysosomes revealed increased heterogeneity in MFM signal from tissues of SCI animals. Ultrastructural analysis using TEM elucidated that both cytosolic and lysosomal ferritin(iron) density was increased in the injured (spinal cord) and non-injured (spleen) tissues of SCI as compared to naïve animals. In-situ EELs analysis revealed that ferritin(iron) was primarily in Fe3+ oxidation state in both naïve and SCI animal tissues. The insights provided by this study and the approaches utilized here can be applied broadly to other systemic problems involving iron regulation or to understand the fate of exogenously delivered iron-oxide nanoparticles.
Asunto(s)
Ferritinas/metabolismo , Hierro/metabolismo , Nanopartículas del Metal/química , Traumatismos de la Médula Espinal/metabolismo , Animales , Citosol/química , Citosol/metabolismo , Citosol/ultraestructura , Modelos Animales de Enfermedad , Ferritinas/química , Ferritinas/ultraestructura , Humanos , Hierro/química , Lisosomas/efectos de los fármacos , Lisosomas/ultraestructura , Nanopartículas del Metal/efectos adversos , Nanopartículas del Metal/uso terapéutico , Microscopía Electrónica de Transmisión , Ratas , Roedores , Espectroscopía de Pérdida de Energía de Electrones , Traumatismos de la Médula Espinal/tratamiento farmacológico , Bazo/química , Bazo/metabolismo , Bazo/ultraestructuraRESUMEN
Organophosphorus flame retardants (PFRs) are extensively used as alternatives to banned polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecane (HBCD). In this study, we analyzed 14 PFRs by means of four mass-spectrometry-based methods: gas chromatography combined with electron-impact mass spectrometry (GC-EI-MS) or negative-chemical-ionization mass spectrometry (GC-NCI-MS) and liquid chromatography combined with tandem mass spectrometry using electrospray ionization (LC-ESI-MS/MS) or atmospheric pressure chemical ionization (LC-APCI-MS/MS). The limits of quantification (LOQs) for LC-ESI-MS/MS and LC-APCI-MS/MS (0.81-970 pg) were 1-2 orders of magnitude lower than the LOQs for GC-EI-MS and GC-NCI-MS (2.3-3900 pg). LC-APCI-MS/MS showed the lowest LOQs (mean = 41 pg; median = 3.4 pg) for all but two of the PFRs targeted in this study. For LC-APCI-MS/MS, the lowest LOQ was observed for tributyl phosphate (TBP) (0.81 pg), and the highest was observed for tris(butoxyethyl) phosphate (TBOEP) (36 pg). The results of this study indicate that LC-APCI-MS/MS is the optimum analytical method for the target PFRs, at least in terms of LOQ.
Asunto(s)
Retardadores de Llama/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Organofosfatos/análisis , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida , Éteres Difenilos Halogenados/análisis , Hidrocarburos Bromados/análisis , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
Metal nanoparticles (NPs) have unique physicochemical properties and a widespread application scope depending on their composition and surface characteristics. Potential biomedical applications and the growing diversity of novel nanocomposites highlight the need for toxicological hazard assessment of next-generation magnetic nanomaterials. Our study aimed to evaluate the cytotoxic and genotoxic properties of coated and uncoated ferric cobalt boron (FeCoB) NPs (5-15 nm particle size) in cultured normal human dermal fibroblasts. Cell proliferation was assessed via ATP bioluminescence kit, and DNA breakage and chromosomal damage were measured by alkaline comet assay and micronucleus test. Polyacryl acid-coated FeCoB NPs [polyacrylic acid (PAA)-FeCoB NPs) and uncoated FeCoB NPs inhibited cell proliferation at 10 µg/ml. DNA strand breaks were significantly increased by PAA-coated FeCoB NPs, uncoated FeCoB NPs and l-cysteine-coated FeCoB NPs (Cys-FeCoB NPs), although high concentrations (10 µg/ml) of coated NPs (Cys- and PAA-FeCoB NPs) showed significantly more DNA breakage when compared to uncoated ones. Uncoated FeCoB NPs and coated NPs (PAA-FeCoB NPs) also induced the formation of micronuclei. Additionally, PAA-coated NPs and uncoated FeCoB NPs showed a negative correlation between cell proliferation and DNA strand breaks, suggesting a common pathomechanism, possibly by oxidation-induced DNA damage. We conclude that uncoated FeCoB NPs are cytotoxic and genotoxic at in vitro conditions. Surface coating of FeCoB NPs with Cys and PAA does not prevent but rather aggravates DNA damage. Further safety assessment and a well-considered choice of surface coating are needed prior to application of FeCoB nanocomposites in biomedicine.
Asunto(s)
Compuestos de Boro/toxicidad , Cobalto/toxicidad , Compuestos Férricos/toxicidad , Campos Magnéticos/efectos adversos , Nanopartículas del Metal/toxicidad , Nanocompuestos/toxicidad , Piel/efectos de los fármacos , Compuestos de Boro/química , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Cobalto/química , Ensayo Cometa , Roturas del ADN/efectos de los fármacos , Contaminantes Ambientales/química , Contaminantes Ambientales/toxicidad , Compuestos Férricos/química , Humanos , Nanopartículas del Metal/química , Nanopartículas del Metal/ultraestructura , Pruebas de Micronúcleos , Microscopía Electrónica de Transmisión , Nanocompuestos/química , Nanocompuestos/ultraestructura , Estrés Oxidativo/efectos de los fármacos , Tamaño de la Partícula , Piel/citología , Espectroscopía de Pérdida de Energía de Electrones , Propiedades de SuperficieRESUMEN
Ceria nanoparticles (NPs) rapidly and easily cycle between Ce(III) and Ce(IV) oxidation states, making them prime candidates for commercial and other applications. Increased commercial use has resulted in increased discharge to the environment and increased associated risk. Once in complex media such as environmental waters or toxicology exposure media, the same redox transformations can occur, causing altered behavior and effects compared to the pristine NPs. This study used high resolution scanning transmission electron microscopy and electron energy loss spectroscopy to investigate changes in structure and oxidation state of small, polymer-coated ceria suspensions in complex media. NPs initially in either the III or IV oxidation states, but otherwise identical, were used. Ce(IV) NPs were changed to mixed (III, IV) NPs at high ionic strengths, while the presence of natural organic macromolecules (NOM) stabilized the oxidation state and increased crystallinity. The Ce(III) NPs remained as Ce(III) at high ionic strengths, but were modified by the presence of NOM, causing reduced crystallinity and degradation of the NPs. Subtle changes to NP properties upon addition to environmental or ecotoxicology media suggest that there may be small but important effects on fate and effects of NPs compared to their pristine form.
Asunto(s)
Cerio , Nanopartículas , Ecotoxicología , Espectroscopía de Pérdida de Energía de Electrones , SuspensionesRESUMEN
This paper addresses a novel approach to atomic resolution elemental mapping, demonstrating a method that produces elemental maps with a similar resolution to the established method of electron energy-loss spectroscopy in scanning transmission electron microscopy. Dubbed energy-filtered imaging scanning transmission electron microscopy (EFISTEM) this mode of imaging is, by the quantum mechanical principle of reciprocity, equivalent to tilting the probe in energy-filtered transmission electron microscopy (EFTEM) through a cone and incoherently averaging the results. In this paper we present a proof-of-principle EFISTEM experimental study on strontium titanate. The present approach, made possible by chromatic aberration correction, has the advantage that it provides elemental maps which are immune to spatial incoherence in the electron source, coherent aberrations in the probe-forming lens and probe jitter. The veracity of the experiment is supported by quantum mechanical image simulations, which provide an insight into the image-forming process. Elemental maps obtained in EFTEM suffer from the effect known as preservation of elastic contrast, which, for example, can lead to a given atomic species appearing to be in atomic columns where it is not to be found. EFISTEM very substantially reduces the preservation of elastic contrast and yields images which show stability of contrast with changing thickness. The experimental application is demonstrated in a proof-of-principle study on strontium titanate.
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Microscopía Electrónica de Transmisión de Rastreo , Energía Filtrada en la Transmisión por Microscopía Electrónica , Simulación por Computador , Electrones , Modelos Teóricos , Óxidos/química , Teoría Cuántica , Espectroscopía de Pérdida de Energía de Electrones , Estroncio/química , Titanio/químicaRESUMEN
Heavy metal pollution has increased in the last decades. Water sources are contaminated and human exposure is often long term exposure to variable amounts of different metals. In this study, male Sprague-Dawley rats were exposed via oral gavage for 28 days to cadmium (Cd) and chromium (Cr), alone and in combination at concentrations 1000 times the human World Health Organization's acceptable water limits. Rat equivalent dosages were used. Blood markers of liver and kidney function were measured, changes to cellular morphology was determined with transmission electron microscopy and the intracellular metal localisation was determined with the electron energy-loss spectroscopy and energy filtered transmission electron microscopy analysis. Both Cd and Cr caused changes to the nuclear and mitochondrial membranes and irregular chromatin condensation of hepatocytes. Cr exposure caused dilation of the rough endoplasmic reticulum (rER). The combination caused nuclear and mitochondrial membrane damage as well as irregular chromatin condensation. In the kidney tissue, Cd caused irregular chromatin condensation in the cells of the proximal convoluted tubule (PCT). Cr caused changes to the outer nuclear and mitochondrial membrane and chromatin structure. The combination group caused membrane damage, irregular chromatin condensation and rER changes in the PCT. All the metal groups showed damage to the endothelial cells and pedicles, but not to the mesangial cells. Cd and Cr bio-accumulation was observed in the nucleus, mitochondria and rER of the liver and kidney and therefore are responsible for the cellular observed damage that can cause functional changes to the tissues and organs.
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Cadmio/toxicidad , Cromo/toxicidad , Riñón/ultraestructura , Hígado/ultraestructura , Animales , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Masculino , Microscopía Electrónica de Transmisión , Ratas , Ratas Sprague-Dawley , Espectroscopía de Pérdida de Energía de ElectronesRESUMEN
The pioneering contributions of Ondrej Krivanek to the development of electron energy loss spectrometers, energy filters, and detectors for transmission and scanning transmission electron microscopes have provided researchers with indispensible tools across a wide range of disciplines in the physical sciences, ranging from condensed matter physics, to chemistry, mineralogy, materials science, and nanotechnology. In addition, the same instrumentation has extended its reach into the life sciences, and it is this aspect of Ondrej Krivanek's influential contributions that will be surveyed here, together with some personal recollections. Traditionally, electron microscopy has given a purely morphological view of the biological structures that compose cells and tissues. However, the availability of high-performance electron energy loss spectrometers and energy filters offers complementary information about the elemental and chemical composition at the subcellular scale. Such information has proven to be valuable for applications in cell and structural biology, microbiology, histology, pathology, and more generally in the biomedical sciences.
Asunto(s)
Disciplinas de las Ciencias Biológicas/instrumentación , Energía Filtrada en la Transmisión por Microscopía Electrónica/métodos , Espectroscopía de Pérdida de Energía de Electrones/métodos , Electrones , Humanos , Nanotecnología/instrumentaciónRESUMEN
Four-dimensional ultrafast electron microscopy (4D-UEM) is a novel analytical technique that aims to fulfill the long-held dream of researchers to investigate materials at extremely short spatial and temporal resolutions by integrating the excellent spatial resolution of electron microscopes with the temporal resolution of ultrafast femtosecond laser-based spectroscopy. The ingenious use of pulsed photoelectrons to probe surfaces and volumes of materials enables time-resolved snapshots of the dynamics to be captured in a way hitherto impossible by other conventional techniques. The flexibility of 4D-UEM lies in the fact that it can be used in both the scanning (S-UEM) and transmission (UEM) modes depending upon the type of electron microscope involved. While UEM can be employed to monitor elementary structural changes and phase transitions in samples using real-space mapping, diffraction, electron energy-loss spectroscopy, and tomography, S-UEM is well suited to map ultrafast dynamical events on materials surfaces in space and time. This review provides an overview of the unique features that distinguish these techniques and also illustrates the applications of both S-UEM and UEM to a multitude of problems relevant to materials science and chemistry.
Asunto(s)
Microscopía Electrónica , Espectroscopía de Pérdida de Energía de Electrones , Análisis EspectralRESUMEN
BACKGROUND: This work is focused on mechanisms of uptake in cancer cells of rationally designed, covalently assembled nanoparticles, made of superparamagnetic iron oxide nanoparticles (SPIONs), fluorophores (doxorubicin or Nile Blue), polyethylene glycol (PEG) and folic acid (FA), referred hereinafter as SFP-FA. METHODS: SFP-FA were characterized by DLS, zetametry and fluorescence spectroscopy. The SFP-FA uptake in cancer cells was monitored using fluorescence-based methods like fluorescence-assisted cell sorting, CLSM with single-photon and two-photon excitation. The SFP-FA endocytosis was also analyzed with electron microscopy approaches: TEM, HAADF-STEM and EELS. RESULTS: The SFP-FA have zeta potential below -6mW and stable hydrodynamic diameter close to 100nm in aqueous suspensions of pH range from 5 to 8. They contain ca. 109 PEG-FA, 480 PEG-OCH3 and 22-27 fluorophore molecules per SPION. The fluorophores protected under the PEG shell allows a reliable detection of intracellular NPs. SFP-FA readily enter into all the cancer cell lines studied and accumulate in lysosomes, mostly via clathrin-dependent endocytosis, whatever the FR status on the cells. CONCLUSIONS: The present study highlights the advantages of rational design of nanosystems as well as the possible involvement of direct molecular interactions of PEG and FA with cellular membranes, not limited to FA-FR recognition, in the mechanisms of their endocytosis. GENERAL SIGNIFICANCE: Composition, magnetic and optical properties of the SFP-FA as well their ability to enter cancer cells are promising for their applications in cancer theranosis. Combination of complementary analytical approaches is relevant to understand the nanoparticles behavior in suspension and in contact with cells.