This minireview describes high-frequency and high-field electron paramagnetic resonance (HFEPR) spectroscopy in the context of its application to bioinorganic chemistry, specifically to metalloproteins and model compounds. HFEPR is defined as frequencies above ~100 GHz (i.e., above W-band) and a resonant field reaching 25 T and above. The ability of HFEPR to provide high-resolution determination of g values of S = 1/2 is shown; however, the main aim of the minireview is to demonstrate how HFEPR can extract spin Hamiltonian parameters [zero-field splitting (zfs) and g values] for species with S > 1/2 with an accuracy and precision unrivalled by other physical methods. Background theory on the nature of zfs in S = 1, 3/2, 2, and 5/2 systems is presented, along with selected examples of HFEPR spectroscopy of each that are relevant to bioinorganic chemistry. The minireview also provides some suggestions of specific systems in bioinorganic chemistry where HFEPR could be rewardingly applied, in the hope of inspiring workers in this area.

High-field, high-frequency electron paramagnetic resonance (EPR) spectroscopy at W-(∼94 GHz) and D-band (∼140 GHz) is important for investigating the conformational dynamics of flexible biological macromolecules because this frequency range has increased spectral sensitivity to nitroxide motion over the 100 ps to 2 ns regime. However, low concentration sensitivity remains a roadblock for studying aqueous samples at high magnetic fields. Here, we examine the sensitivity of a non-resonant thin-layer cylindrical sample holder, coupled to a quasi-optical induction-mode W-band EPR spectrometer (HiPER), for continuous wave (CW) EPR analyses of: (i) the aqueous nitroxide standard, TEMPO; (ii) the unstructured to α-helical transition of a model IDP protein; and (iii) the base-stacking transition in a kink-turn motif of a large 232 nt RNA. For sample volumes of ∼50 μL, concentration sensitivities of 2-20 μM were achieved, representing a ∼10-fold enhancement compared to a cylindrical TE011 resonator on a commercial Bruker W-band spectrometer. These results therefore highlight the sensitivity of the thin-layer sample holders employed in HiPER for spin-labeling studies of biological macromolecules at high fields, where applications can extend to other systems that are facilitated by the modest sample volumes and ease of sample loading and geometry.Copyright © 2016 Elsevier Inc. All rights reserved.

We describe a novel cw millimeter-wave electron spin resonance (ESR) spectrometer designed to operate in the frequency range of 80–200 GHz and in the temperature range of 2.5–300 K, which may be easily scaled to higher frequencies. The spectrometer uses a bimodal reflection cavity coupled to a circular corrugated guide and uses Gaussian quasi-optics for most of the front-end signal processing. This technique has very low insertion loss and allows a number of sophisticated measurement techniques to be employed including induction operation, which significantly reduces the effect of microphonics and stray reflections. A number of examples are given illustrating the sensitivity of the instrument and the advantages of using ESR at high fields.

Magnetic resonance is a very powerful methodology that has been employed successfully in many applications for about 70years now, resulting in a wealth of scientific, technological, and diagnostic data. Despite its many advantages, one major drawback of magnetic resonance is its relatively poor sensitivity and, as a consequence, its bad spatial resolution when examining heterogeneous samples. Contemporary science and technology often make use of very small amounts of material and examine heterogeneity on a very small length scale, both of which are well beyond the current capabilities of conventional magnetic resonance. It is therefore very important to significantly improve both the sensitivity and the spatial resolution of magnetic resonance techniques. The quest for higher sensitivity led in recent years to the development of many alternative detection techniques that seem to rival and challenge the conventional "old-fashioned" induction-detection approach. The aim of this manuscript is to briefly review recent advances in the field, and to provide a quantitative as well as qualitative comparison between various detection methods with an eye to future potential advances and developments. We first offer a common definition of sensitivity in magnetic resonance to enable proper quantitative comparisons between various detection methods. Following that, up-to-date information about the sensitivity capabilities of the leading recently-developed detection approaches in magnetic resonance is provided, accompanied by a critical comparison between them and induction detection. Our conclusion from this comparison is that induction detection is still indispensable, and as such, it is very important to look for ways to significantly improve it. To do so, we provide expressions for the sensitivity of induction-detection, derived from both classical and quantum mechanics, that identify its main limiting factors. Examples from current literature, as well as a description of new ideas, show how these limiting factors can be mitigated to significantly improve the sensitivity of induction detection. Finally, we outline some directions for the possible applications of high-sensitivity induction detection in the field of electron spin resonance.Copyright © 2017 Elsevier Inc. All rights reserved.

Quantum information, encoded within the states of quantum systems, represents a novel and rich form of information which has inspired new types of computers and communications systems. Many diverse electron spin systems have been studied with a view to storing quantum information, including molecular radicals, point defects and impurities in inorganic systems, and quantum dots in semiconductor devices. In these systems, spin coherence times can exceed seconds, single spins can be addressed through electrical and optical methods, and new spin systems with advantageous properties continue to be identified. Spin ensembles strongly coupled to microwave resonators can, in principle, be used to store the coherent states of single microwave photons, enabling so-called microwave quantum memories. We discuss key requirements in realising such memories, including considerations for superconducting resonators whose frequency can be tuned onto resonance with the spins. Finally, progress towards microwave quantum memories and other developments in the field of superconducting quantum devices are being used to push the limits of sensitivity of inductively-detected electron spin resonance. The state-of-the-art currently stands at around 65 spins per Hz, with prospects to scale down to even fewer spins.Copyright © 2017. Published by Elsevier Inc.

High-field/high-frequency electron spin resonance (ESR) offers improved sensitivity and resolution compared to ESR at conventional fields and frequencies. However, most high-field/high-frequency ESR spectrometers suffer from limited mm-wave power, thereby requiring long mm-wave pulses. This precludes their use when relaxation times are short, e.g., in fluid samples. Low mm-wave power is also a major factor limiting the achievable spectral coverage and thereby the multiplex advantage of Fourier transform ESR (FTESR) experiments. High-power pulses are needed to perform two-dimensional (2D) FTESR experiments, which can unravel the dynamics of a spin system in great detail, making it an excellent tool for studying spin and molecular dynamics. We report on the design and implementation of a high-power, high-bandwidth, pulsed ESR spectrometer operating at 95 GHz. One of the principal design goals was the ability to investigate dynamic processes in aqueous samples at physiological temperatures with the intent to study biological systems. In initial experiments on aqueous samples at room temperature, we achieved 200 MHz spectral coverage at a sensitivity of 1.1×1010s spins and a dead time of less than 50 ns. 2D-electron-electron double resonance experiments on aqueous samples are discussed to demonstrate the practical application of such a spectrometer.

We describe a quasioptical 94 GHz kW pulsed electron paramagnetic resonance spectrometer featuring π/2 pulses as short as 5 ns and an instantaneous bandwidth of 1 GHz in nonresonant sample holders operating in induction mode and at low temperatures. Low power pulses can be as short as 200 ps and kilowatt pulses as short as 1.5 ns with timing resolution of a few hundred picoseconds. Phase and frequency can be changed on nanosecond time scales and complex high power pulse sequences can be run at repetition rates up to 80 kHz with low dead time. We demonstrate that the combination of high power pulses at high frequencies and nonresonant cavities can offer excellent concentration sensitivity for orientation selective pulsed electron double resonance (double electron-electron resonance), where we demonstrate measurements at 1 μM concentration levels.

We provide a review of current electron spin resonance (ESR) techniques for studying basic molecular mechanisms in membranes and proteins by using nitroxide spin labels. In particular, nitroxide spin label studies with high-field/high-frequency ESR and two-dimensional Fourier transform ESR enable one to accurately determine distances in biomolecules, unravel the details of the complex dynamics in proteins, characterize the dynamic structure of membrane domains, and discriminate between bulk lipids and boundary lipids that coat transmembrane peptides or proteins; these studies can also provide time resolution to studies of functional dynamics of proteins. We illustrate these capabilities with recent examples.

Complexity of paramagnetic catalysts and materials increases, and the same applies to systems targeted by integrative structural biology. Hence, EPR spectroscopists must find ways to enhance information content of their data. I argue that a third major wave of method development in EPR spectroscopy, which is triggered by recent advances in digital electronics and computing, can achieve this. Transfer of NMR methods to EPR will go on, but part of the new EPR methodology will depend on completely new concepts.Copyright © 2019 Elsevier Inc. All rights reserved.

This minireview describes high-frequency and high-field electron paramagnetic resonance (HFEPR) spectroscopy in the context of its application to bioinorganic chemistry, specifically to metalloproteins and model compounds. HFEPR is defined as frequencies above ~100 GHz (i.e., above W-band) and a resonant field reaching 25 T and above. The ability of HFEPR to provide high-resolution determination of g values of S = 1/2 is shown; however, the main aim of the minireview is to demonstrate how HFEPR can extract spin Hamiltonian parameters [zero-field splitting (zfs) and g values] for species with S > 1/2 with an accuracy and precision unrivalled by other physical methods. Background theory on the nature of zfs in S = 1, 3/2, 2, and 5/2 systems is presented, along with selected examples of HFEPR spectroscopy of each that are relevant to bioinorganic chemistry. The minireview also provides some suggestions of specific systems in bioinorganic chemistry where HFEPR could be rewardingly applied, in the hope of inspiring workers in this area.

We describe a tunable electron paramagnetic resonance (EPR) spectrometer designed to operate at frequencies between 160 and 525 GHz and magnetic fields of up to 20 T. To operate in such a broad frequency range we use a very stable optically pumped far infrared laser. The performance of the spectrometer has been measured with solid and liquid samples. This allows us to outline the potential uses of the spectrometer.

We present design details of and first measurements with a novel continuous wave (cw) high-field/high-frequency electron paramagnetic resonance spectrometer operating at a microwave frequency of 360 GHz and a magnetic field of up to 14 T. The spectrometer design incorporates a heterodyne mixer detection scheme with a quasi-optical transmission line and a bimodal induction mode Fabry–Perot cavity. First cw experiments on polycrystalline 1,1-diphenyl-2-picryl-hydrazyl and bisdiphenylene-β-phenylallyl benzolate in polystyrene at room temperature and 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl in frozen solution at 190 K demonstrate the high Zeeman resolution achievable and allow an estimate of the present detection sensitivity of 4×109 spins/G at a detection bandwidth of 1 Hz.

We describe a novel cw millimeter-wave electron spin resonance (ESR) spectrometer designed to operate in the frequency range of 80–200 GHz and in the temperature range of 2.5–300 K, which may be easily scaled to higher frequencies. The spectrometer uses a bimodal reflection cavity coupled to a circular corrugated guide and uses Gaussian quasi-optics for most of the front-end signal processing. This technique has very low insertion loss and allows a number of sophisticated measurement techniques to be employed including induction operation, which significantly reduces the effect of microphonics and stray reflections. A number of examples are given illustrating the sensitivity of the instrument and the advantages of using ESR at high fields.

We report methodology that combines an ultrawide band multifrequency microwave system with technology of high magnetic fields for solving challenging problems in electron magnetic resonance (EMR) spectroscopy. This strategy has been made possible due to a novel EMR facility operating in an exceptionally wide range of microwave frequencies of 24 GHz to 3 THz, at magnetic fields up to 17 T, and in the temperature range of 1.6 to 330 K. The basic configuration of the multifrequency system works in a transmission mode and employs oversized cylindrical waveguides for routing the microwave power. A wide-band, low-noise, liquid helium cooled (4.2 K) InSb bolometer is used for signal detection. This approach results in an extremely wide-band performance, thus making it possible to employ a variety of solid-state millimeter and submillimeter microwave sources in combination with a far infrared laser microwave source for performing multifrequency EMR experiments. A complexity of resonant structures and related technical problems such as microphonics at high magnetic fields is virtually eliminated. The system is simple, yet sensitive, and has been revealed to be extremely advantageous while solving such problems as observation of AFMR transitions in spin-ordered systems, g-factor resolution enhancement in complex organic radicals, and resonance signal detection in EMR-silent spin systems having integer spin and large zero field splitting. A technical description of the multifrequency high-field EMR facility is presented and results of its performance tests are given. The potential utility of using the multifrequency high-field methodology in EMR studies is illustrated with selected examples of its recent applications.Copyright 2000 Academic Press.

An electron paramagnetic resonance (EPR) spectrometer is described which allows for continuous-wave and pulsed EPR experiments at 275 GHz (wavelength 1.1 mm). The related magnetic field of 9.9 T for g approximately 2 is supplied by a superconducting solenoid. The microwave bridge employs quasi-optical as well as conventional waveguide components. A cylindrical, single-mode cavity provides a high filling factor and a high sensitivity for EPR detection. Even with the available microwave power of 1 mW incident at the cavity a high microwave magnetic field B1 is obtained of about 0.1 mT which permits pi/2-pulses as short as 100 ns. The performance of the spectrometer is illustrated with the help of spectra taken with several samples.

Electron paramagnetic resonance (EPR) and double electron-electron resonance (DEER) spectroscopies are powerful and versatile tools for studying local structures and dynamic properties of biological molecules. Similar to nuclear magnetic resonance (NMR) spectroscopy, EPR/DEER spectroscopies become more advantageous at higher frequencies and higher magnetic fields because of better spectral resolution as well as higher spin polarization. Here, we describe development of a high-frequency (HF) EPR/DEER spectrometer operating in the frequency range of 107-120 and 215-240 GHz and in the magnetic field range of 0-12.1 T, which has unique experimental capabilities such as enabling the complete spin polarization and wide-band DEER spectroscopy. Emphasis is given on the application of HF EPR/DEER techniques, and specific examples of HF EPR spectroscopy to drastically increase spin coherence in nanodiamonds as well as HF DEER spectroscopy to extract spin concentration in a diamond crystal are presented.© 2015 Elsevier Inc. All rights reserved.

An electron-spin-echo-detected, electron-paramagnetic-resonance study has been performed on the type 2 copper site of quercetin 2,3-dioxygenase from Aspergillus japonicus. In the protein, copper is coordinated by three histidine nitrogens and two sulfurs from the inhibitor diethyldithiocarbamate. A single crystal of the protein was studied at 95 GHz and the complete g-tensor determined. The electron-paramagnetic-resonance data are compatible with two orientations of the principal g-axes in the copper center, one of which is preferred on the basis of an analysis of the copper coordination and the d-orbitals that are involved in the unpaired-electron orbital. For this orientation, the principal z-axis of the g-tensor makes an angle of 19 degrees with the Cu-N(His112) bond and the N of His112 may be considered the axial ligand. The singly occupied molecular orbital contains a linear combination of copper dxy and dyz-orbitals, which are antibonding with atomic orbitals of histidine nitrogens and diethyldithiocarbamate sulfurs. The orientation of the g-tensor for the quercetin 2,3-dioxygenase is compared with that for type 1 copper sites.

Tryptophan-based free radicals have been implicated in a myriad of catalytic and electron transfer reactions in biology. However, very few of them have been trapped so that biophysical characterizations can be performed in a high-precision context. In this work, tryptophan derivative-based radicals were studied by high-frequency/high-field electron paramagnetic resonance (HFEPR) and quantum chemical calculations. Radicals were generated at liquid nitrogen temperature with a photocatalyst, sacrificial oxidant, and violet laser. The precise g-anisotropies of l- and d-tryptophan, 5-hydroxytryptophan, 5-methoxytryptophan, 5-fluorotryptophan, and 7-hydroxytryptophan were measured directly by HFEPR. Quantum chemical calculations were conducted to predict both neutral and cationic radical spectra for comparison with the experimental data. The results indicate that under the experimental conditions, all radicals formed were cationic. Spin densities of the radicals were also calculated. The various line patterns and g-anisotropies observed by HFEPR can be understood in terms of spin-density populations and the positioning of oxygen atom substitution on the tryptophan ring. The results are considered in the light of the tryptophan and 7-hydroxytryptophan diradical found in the biosynthesis of the tryptophan tryptophylquinone cofactor of methylamine dehydrogenase.

Electron paramagnetic resonance (EPR) spectroscopy (also known as electron spin resonance, ESR, or electron magnetic resonance, EMR, spectroscopy) is often described as the "gold standard" for the detection and characterisation of radicals in chemical, biological and medical systems. The article reviews aspects of EPR spectroscopy and discusses how this methodology and related techniques can be used to obtain useful information from biological systems. Consideration is given to the direct detection of radicals, the use of spin traps and the detection of nitric oxide, and the advantages and pitfalls of various approaches. When used with care, this technique can provide a huge amount of valuable data on the presence of radicals, their identity and information on their concentration, structure, mobility and interactions. It is however a technique that has limitations, and the novice user needs to understand the various pitfalls and shortcomings of the method to avoid making significant errors.Copyright © 2016 Elsevier Inc. All rights reserved.

Coordination complexes that possess large magnetic anisotropy (otherwise known as zero-field splitting, ZFS) have possible applications in the field of magnetic materials, including single molecule magnets (SMMs). Previous studies have explored the role of coordination number and geometry in controlling the magnetic anisotropy and SMM behavior of high-spin ( = 3/2) Co(II) complexes. Building upon these efforts, the present work examines the impact of ligand oxidation state and structural distortions on the spin states and ZFS parameters of pentacoordinate Co(II) complexes. The five complexes included in this study (-) have the general formula, [Co(Tp)()] (X = O, S; Y = N, O; = 0 or 1), where Tp is the scorpionate ligand hydrotris(3,5-diphenyl-pyrazolyl)borate(1-) and are bidentate dioxolene-type ligands that can access multiple oxidation states. The specific ligands used herein are 4,6-di--butyl substituted -aminophenolate and -aminothiophenolate ( and, respectively), -iminosemiquinonate and -semiquinonate radicals ( and, respectively), and -iminobenzoquinone (). Each complex exhibits a distorted trigonal bipyramidal geometry, as revealed by single-crystal X-ray diffraction. Direct current (dc) magnetic susceptibility experiments confirmed that the complexes with closed-shell ligands (,, and ) possess = 3/2 ground states with negative -values (easy-axis anisotropy) of -41, -78, and -30 cm, respectively. For and, antiferromagnetic coupling between the Co(II) center and -(imino)semiquinonate radical ligand results in = 1 ground states that likewise exhibit very large and negative anisotropy (-100 >> -140 cm). Notably, ZFS was measured directly for each complex using far-infrared magnetic spectroscopy (FIRMS). In combination with high-frequency and -field electron paramagnetic resonance (HFEPR) studies, these techniques provided precise spin-Hamiltonian parameters for complexes,, and. Multireference calculations, using the CASSCF/NEVPT2 approach, indicate that the strongly negative anisotropies of these Co(II) complexes arise primarily from distortions in the equatorial plane due to constrictions imposed by the Tp ligand. This effect is further amplified by cobalt(II)-radical exchange interactions in and.

High-field/high-frequency electron paramagnetic resonance spectroscopy was employed to synthesize melanin to provide relevant information on the carbon-centered free-radical nature of this biomaterial.