FID detection of EPR and ENDOR spectra at high microwave frequencies

High-frequency pulsed EPR spectroscopy allows FID detection of EPR spectra owing to the short dead time that can be achieved. This FID detection is particularly attractive for EPR and ENDOR spectroscopy of paramagnetic species that exhibit inhomogeneously broadened EPR lines and short dephasing times. Experiments are reported for the metalloprotein azurin at 275 GHz.     

Probing Protein Secondary Structure Using EPR: Investigating a Dynamic Region of Visual Arrestin

One key application of site-directed spin labeling electron paramagnetic resonance (EPR) spectroscopy is the determination of sequence-specific secondary structure in proteins. Regular secondary structure leads to a periodic variation in both side chain motion and solvent accessibility, two properties easily monitored by EPR techniques. Specifically, saturation recovery (SR) EPR spectroscopy has proven to be useful for making accessibility measurements for multiple protein structure populations by determining individual accessibilities and is, therefore, well suited to study the structure of proteins exhibiting multiple conformations in equilibrium. Here we employ both continuous wave and SR EPR spectroscopy in combination to examine the secondary structure of a short sequence showing conformational heterogeneity in visual rod arrestin. The EPR data presented here clearly distinguish between the unstructured loop and the helical structure formed in the crystallographic tetramer of visual arrestin and show that this region is unstructured in solution.     

Millimeter and submillimeter EPR spectroscopy

For a long time, the electron paramagnetic resonance (EPR) spectrometers have been operated in X and Q bands with wavelengths of microwave radiation about 3 cm and 8 mm. Increasing the operating frequency improves the basic parameters of an EPR spectrometer. In view of this, there has recently been rapid development of high-frequency EPR spectroscopy, including the submillimeter-wave range, related to significant progress in the millimeter-and submillimeterwave technique. This paper discusses characteristic features, application areas, and the state of the art of the experimental technique of EPR spectroscopy in the millimeter-wave range and the short-wavelength region of the millimeter-wave range. The design features of the high-frequency EPR spectrometer operated in the frequency range 65–1500 GHz, which was created at the E. K. Zavoisky Physico-Technical Institute of the Kazan’ Scientific Center of the Russian Academy of Sciences, are presented. The results of studying the structure of the paramagnetic centers formed by impurity Ho³⁺ ions in synthetic forsterite (Mg₂SiO₄), obtained by the method of tunable high-frequency EPR spectroscopy, are reported.     

Detection of glass foreign bodies in soft tissues with low-frequency EPR spectroscopy

The detection of glass particles in glass-induced injuries is often difficult. Most commercial glasses contain iron (Fe³⁺) centers which present a characteristic electron paramagnetic resonance (EPR) spectrum nearg = 4.3. The detection of small glass particles in vivo can be achieved with low-frequency EPR spectroscopy (1.2 GHz) operating at low field. The method was sucessfully applied in anesthetized live animals where small pieces of glass were implanted under the skin in their backs. This could be a new clinically relevant application of EPR spectroscopy.     

Paramagnetic centers in tumor cells

Free radical properties of different types of tumor cells were compared. Electron paramagnetic resonance (EPR) studies were performed for human (BM, IGR and SK) and mouse (B16 and S91) melanoma cells. In contrast to melanotic melanoma IGR, BM and B16 cells, amelanotic S91 cells contained only a trace amount of melanin. No EPR signals were detected for Caco2 cells and only a very weak EPR line was measured for fibroblast cells. Melanin does not exist in these cells. The aim of this work was the application of EPR spectroscopy to the determination of the kind of melanin (eu- or pheomelanin) in melanotic tumor cells. Microwave saturation of EPR spectra of tumor cells with high and low melanin content was compared. Eumelanin was identified in human BM, IGR, SK, and B16 melanoma cells. Single asymmetrical EPR lines were detected for these samples. The EPR spectra of human BM melanoma cells had the highest intensity. Paramagnetic centers in amelanotic S91 melanoma cells were also found. Trace amounts of eumelanin free radicals and the other free radicals in cells were responsible for their very weak EPR lines. The obtained results indicate that EPR spectroscopy is a very useful technique for the identification of melanin in tumor cells. Strong differences of microwave saturation of EPR lines for cells with high and low melanin content were observed. EPR lines of tumor cells with a low melanin content did not saturate at the used range of microwave power. Saturation was observed for melanotic BM melanoma cells.     

Nonequivalent spectra of unpaired electrons in field and frequency modulation

We report a difference in the spectral lineshapes of continuous-wave (CW) electron paramagnetic resonance (EPR) spectroscopy between field and frequency modulation. This finding addresses the long-standing question of the effect of modulation in EPR absorption. We compared the first-derivative EPR spectra at 1.1 GHz for lithium phthalocyanine crystals, which have a single narrow linewidth in the EPR absorption spectrum, using field and frequency modulation. The experimental findings suggest that unpaired electrons have different behaviors under perturbation due to field and frequency modulation.     

Effect of nitrogen dioxide on the EPR property of lithium octa-n-butoxy 2,3-naphthalocyanine (LiNc-BuO) microcrystals

Lithium octa-n-butoxy-naphthalocyanine (LiNc-BuO) is a stable free radical that can be detected by electron paramagnetic resonance (EPR) spectroscopy. Previously we have reported that microcrystals of LiNc-BuO exhibit a single sharp EPR peak, whose width varies linearly with the partial pressure of paramagnetic molecules such as oxygen and nitric oxide. In this report, we present the effect of nitrogen dioxide (NO2), which is also a paramagnetic molecule, on the EPR properties of LiNc-BuO. The gas-sensing property of LiNc-BuO is attributed to the open molecular framework of the crystal structure which is arranged with wide channels capable of accommodating large molecules such as NO2. The EPR linewidth of LiNc-BuO was highly sensitive to the partial pressure of NO2 in the gas mixture. The line-broadening was quick and reversible in the short-term for low concentration of NO2. However, the EPR signal intensity decreased with time of exposure, apparently due to a reaction of NO2 with LiNc-BuO crystals to give diamagnetic products. The results suggested that LiNc-BuO may be a useful probe for the determination of trace amounts of NO2 using EPR spectroscopy.     

Aspects of detection of irradiated foods by EPR spectroscopy

Some methodological aspects important for the detection of irradiated foods by the EPR spectroscopy are discussed. These are: specificity and stability of the EPR signals produced by ionizing radiation in foods, complexity of radiation induced EPR signals and their detection at different doses, the dose dependence of the intensities of the EPR signals involved. The importance of the native EPR signals observed in foods for the detection of irradiation and the role of quantitative measurements in the development of detection methods are shown.     

The effects of cholesterol on magnetically aligned phospholipid bilayers: a solid-state NMR and EPR spectroscopy study

This paper presents the first time that both solid-state NMR spectroscopy and EPR spectroscopy are used to study the effects of cholesterol on magnetically aligned phospholipid bilayers (bicelles). Solid-state deuterium NMR spectroscopy was carried out using both chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC-d(54)) and a partially deuterated beta-[2,2,3,4,4,6-(2)H(6)]cholesterol (cholesterol-d(6)). Also, EPR spectroscopy was carried out utilizing a 3 beta-doxyl-5 alpha-cholestane (cholestane) spin probe incorporated into magnetically aligned bilayers to provide a more complete picture about the ordering and dynamics of the phospholipid and cholesterol molecules in the bicelle membrane system. The results demonstrate that cholesterol was successfully incorporated into the phospholipid bilayers. The molecular order parameters extracted directly from the (2)H NMR spectra of both DMPC-d(54) and cholesterol-d(6) were compared to that from the EPR study of cholestane. The order parameters indicate that the sterol was motionally restricted, and that the DMPC had high order and low motion for the hydrocarbon segments close to the head groups of the phospholipids and less order and more rapid motion toward the terminal methyl groups. Both methods clearly indicate an overall increase in the degree of ordering of the molecules in the presence of cholesterol and a decrease in the degree of ordering at higher temperatures. However, EPR spectroscopy and (2)H NMR spectroscopy exhibit different degrees of sensitivity in detecting the phospholipid molecular motions in the membrane. Finally, cholesterol increases the minimum alignment temperature necessary to magnetically align the phospholipid bilayers.     

中国电子顺磁共振(EPR)发展50年回顾

自从1945年前苏联科学家Zavoisky在固体中观察到电子顺磁共振（EPR）这一奇妙的物理现象以来,电子顺磁共振波谱学已经经历了60多年漫长的发展历程. 在世界上,20世纪50、60年代是EPR的基本理论、实验技术和实验方法迅速发展的时期,同时对我国产生了很大的影响. 从60年代开始至今,EPR波谱学在我国已有50多年的发展,其间大约可以分为3个阶段： (一) 20世纪60~80年代初是EPR波谱设备研制和技术发展的初始阶段;(二) 20世纪80年代到21世纪初主要是EPR实验技术进一步发展和实验方法及其在化学、物理学、生物学、医学等学科中的应用研究阶段;(三) 21世纪以来,开始了新的实验技术和开展更加广泛深入的应用研究,特别是开始脉冲EPR波谱学的应用和相关设备的研制.     

Assessment of liver phagocytic activity using EPR spectrometry and imaging

The aim of the present study was to evaluate the usefulness of electron paramagnetic resonance (EPR) spectroscopy and imaging in assessing the phagocytic activity of the liver after administration of India ink. We conducted experiments on livers from control rodents and from rodents in which the Kupffer cell population had been depleted by pretreatment with gadolinium chloride. The EPR signal intensity recorded in liver homogenates was about two times lower in GdCl(3) treated rats than in control rats. EPR imaging carried out on precision-cut liver slices indicated a good correlation between the depletion of Kupffer cells and the EPR signal intensity.     

在体电子自旋共振(EPR)波谱和成像技术在生物医学中的应用

活性氧和氮自由基(ROS/RNS)在一系列的人类疾病中扮演着双重角色. 它们可以是氧化剂, 诱导氧化状态, 导致组织损伤. 它们又可以是信号传导因子, 诱发保护性反应, 使得被调节的组织器官经受得起更强的损伤. 鉴于它们在生物医学中的重要作用, 检测它们产生和分布的技术的研究因而变得必要和紧迫. 在体电子自旋共振(EPR)波谱和成像技术渐已被应用于活体生物体系中用以表针和显像ROS/RNS. EPR 波谱特性(包括线宽、强度和寿命)以及空间分布信息已为动物甚至人体病理模型中氧化还原状态和氧分布的检测提供不可缺少的依据. 该文将简单描述和讨论一系列在体EPR 波谱和成像技术在器官和组织中的应用, 其中包括活体组织氧化还原状态, 活体组织氧分布和时间演化, 自由基空间以及谱-空间成像等.     

Estimation of mean and median pO2 values for a composite EPR spectrum

Electron paramagnetic resonance (EPR)-based oximetry is capable of quantifying oxygen content in samples. However, for a heterogeneous environment with multiple pO2 values, peak-to-peak linewidth of the composite EPR lineshape does not provide a reliable estimate of the overall pO2 in the sample. The estimate, depending on the heterogeneity, can be severely biased towards narrow components. To address this issue, we suggest a postprocessing method to recover the linewidth histogram which can be used in estimating meaningful parameters, such as the mean and median pO2 values. This information, although not as comprehensive as obtained by EPR spectral-spatial imaging, goes beyond what can be generally achieved with conventional EPR spectroscopy. Substantially shorter acquisition times, in comparison to EPR imaging, may prompt its use in clinically relevant models. For validation, simulation and EPR experiment data are presented.     

Mechanism of oxygen response in carbon-based sensors

The mechanism of oxygen response in several newly synthesized oxygen-sensitive chars was studied with the use of EPR spectroscopy. The results suggest that the compounds contain two basic types of paramagnetic centers (PC). The change in oxygen concentration leads to a mutual and reversible transformation of PCs in chars, which is reflected in EPR parameters. The adsorbed molecular oxygen progressively disturbs the wave functions of the PCs and so breaks the Heisenberg exchange between them. At high oxygen concentration, the 2D dipole-dipole interaction between PCs at the surface comes into play and determines the EPR lineshape. A suggested model quantitatively describes the evolution of the basic EPR parameters of each PC as a function of oxygen concentration.     

Pulsed EPR spectrometer with injection-locked UCSB free-electron laser

We describe the first free-electron laser (FEL)-based pulsed electron paramagnetic resonance (EPR) system designed to study spin dynamics and structure changes of proteins in aqueous solution with nano-second of time resolution. This novel approach opens up the possibility for high-power sub-THz and THz pulsed EPR spectroscopy.     

Conformational dynamics of some short-chain biradicals in solutions

Intramolecular electron spin exchange, as a function of temperature and the solvent nature, has been studied by X-band electron paramagnetic resonance (EPR) spectroscopy in five short-chain flexible nitroxide biradicals. Certain thermodynamic parameters of the conformational rearrangements were calculated from the EPR spectra. The process of spin exchange in short flexible biradicals has some peculiarities in comparison with that in long-chain molecules.     

Microstructure of LaCoO3 prepared by freeze-drying of metal-citrate precursors revealed by EPR

Two methods for the preparation of LaCoO₃ samples were used: thermal decomposition of La-Co citrate precursors obtained by freeze-drying of the corresponding solutions and by a solid state reaction. Microstructural characterization was made by electron paramagnetic resonance spectroscopy (EPR). For assignment of the EPR signals, La_1−xSr_xCoO₃ samples were used as EPR references. The LaCoO₃ oxides prepared from citrates and by a solid state reaction were shown to differ in respect of the mean oxidation state of the cobalt ions, the specific surface area and the particle morphology. EPR spectroscopy reveals for ex-citrate LaCoO₃ ferromagnetic Co³⁺ and Co⁴⁺ coupled ions. For LaCoO₃ samples obtained by a solid state reaction, EPR permits detecting Co₃O₄ impurities only.     

The Role of Metal Ions in the Study of Ancient Paper by Electron Paramagnetic Resonance

Samples from covers and endleaves of sixteenth to eighteenth century books in good conservation state from the Biblioteca Nazionale Marciana in Venice have been investigated by means of electron paramagnetic resonance (EPR) spectroscopy and optical microscopy to characterize the embedded paramagnetic species and to test differences and similarities of the materials in the two types of book components. We detected the Mn(II), Fe(III), and Cu(II) paramagnetic ions, and analyzed their EPR signals by comparing them with previous results in literature of EPR studies on paper. Interestingly, the Mn(II) EPR spectrum profile appears as a fingerprint for samples coming from the same book, either from cover or endleaf, and it looks different for samples extracted from different books. We discuss the role of EPR as a spectroscopic tool for characterizing the interaction of the ions with the paper components and as possible agents of degradation.     

Free radical EPR spectroscopy analysis using blind source separation

In this paper, we propose a novel approach for electron paramagnetic resonance (EPR) mixture spectra analysis based on blind source separation (BSS) technique. EPR spectrum of a free radical is often superimposed by overlapping spectra of other species. It is important and challenging to accurately identify and quantify the 'pure' spectra from such mixtures. In this study, an automated BSS method implementing independent component analysis is used to extract the components from mixed EPR spectra that contain overlapping components of different paramagnetic centers. To apply this method, there is no requirement to know the component spectra or the number of components in advance. The method is applied to analyze free radical EPR spectra which are collected from standard chemical system, cultured cell suspense, and ex vivo rat kidneys by spin trapping EPR technique. Results show that the BSS method proposed here is capable of identifying the component EPR spectra from mixtures with unknown compositions. The BSS technique can offer powerful aids in resolving spectral overlapping problems in general EPR spectroscopy analysis.     

Comparison of Continuous Wave, Spin Echo, and Rapid Scan EPR of Irradiated Fused Quartz

The E' defect in irradiated fused quartz has spin lattice relaxation times (T(1)) about 100 to 300 μs and spin-spin relaxation times (T(2)) up to about 200 μs, depending on the concentration of defects and other species in the sample. These long relaxation times make it difficult to record an unsaturated continuous wave (CW) electron paramagnetic resonance (EPR) signal that is free of passage effects. Signals measured at X-band (~9.5 GHz) by three EPR methods: conventional slow-scan field modulated EPR, rapid scan EPR, and pulsed EPR, were compared. To acquire spectra with comparable signal-to-noise, both pulsed and rapid scan EPR require less time than conventional CW EPR. Rapid scan spectroscopy does not require the high power amplifiers that are needed for pulsed EPR. The pulsed spectra, and rapid scan spectra obtained by deconvolution of the experimental data, are free of passage effects.