Continuous Wave Electron Paramagnetic Resonance Research Articles

Rational Design of an Air-Stable, High-Spin Diradical with Diazapyrene.

Stable carbon-based polyradicals exhibiting strong spin-spin coupling and slow depolarization processes are particularly attractive functional materials. A new molecular motif synthesized by a convenient method that allows the integration of stable, high-spin radicals to (hetero)aromatic polycycles has been developed, as illustrated by a non-Kekulé diradical showing a triplet ground state with long persistency (τ1/2 ≈31 h) in air. Compared to the widely used 1,3-phenylene, the newly designed (diaza)pyrene-4,10-diyl moiety is for the first time demonstrated to confer ferromagnetic (FM) spin coupling, allowing delocalized non-disjoint SOMOs. With the X-ray crystallography unambiguously proving the diradical structure, the triplet ground state was thoroughly characterized. A large ΔES-T of 1.1 kcal/mol, proving the strong FM coupling effect, was revealed consistently by superconducting quantum interference device (SQUID) measurements and variable-temperature electron paramagnetic resonance (EPR) spectroscopy, while the zero-field splitting and triplet nutation characters were examined by continuous-wave and pulsed EPR spectroscopy. A millisecond spin-lattice relaxation time was also detected. The current study not only offers a new molecular motif enabling FM coupling between carbon-based spins, but more importantly presents a general method for installing stable polyradicals into functional π-systems.

A Heterometallic Porphyrin Dimer as a Potential Quantum Gate: Magneto‐Structural Correlations and Spin Coherence Properties

AbstractIn the development of two‐qubit quantum gates, precise control over the intramolecular spin‐spin interaction between molecular spin units plays a pivotal role. A weak but measurable exchange coupling is especially important for achieving selective spin addressability that allows controlled manipulation of the computational basis states |00⟩ |01⟩ |10⟩ |11⟩ by microwave pulses. Here, we report the synthesis and Electron Paramagnetic Resonance (EPR) study of a heterometallic meso‐meso (m‐m) singly‐linked VIVO−CuII porphyrin dimer. X‐band continuous wave EPR measurements in frozen solutions suggest a ferromagnetic exchange coupling of ca. 8 ⋅ 10−3 cm−1. This estimation is supported by Density Functional Theory calculations, which also allow disentangling the ferro‐ and antiferromagnetic contributions to the exchange. Pulsed EPR experiments show that the dimer maintains relaxation times similar to the monometallic CuII porphyrins. The addressability of the two individual spins is made possible by the different g‐tensors of VIV and CuII‐ions, in contrast to homometallic dimers where tilting of the porphyrin planes plays a key role. Therefore, single‐spin addressability in the heterometallic dimer can be maintained even with small tilting angles, as expected when deposited on surface, unlocking the full potential of molecular quantum gates for practical applications.

A Heterometallic Porphyrin Dimer as a Potential Quantum Gate: Magneto-Structural Correlations and Spin Coherence Properties.

In the development of two-qubit quantum gates, precise control over the intramolecular spin-spin interaction between molecular spin units plays a pivotal role. A weak but measurable exchange coupling is especially important for achieving selective spin addressability that allows controlled manipulation of the computational basis states |00⟩ |01⟩ |10⟩ |11⟩ by microwave pulses. Here, we report the synthesis and Electron Paramagnetic Resonance (EPR) study of a heterometallic meso-meso (m-m) singly-linked VIV O-CuII porphyrin dimer. X-band continuous wave EPR measurements in frozen solutions suggest a ferromagnetic exchange coupling of ca. 8 ⋅ 10-3 cm-1 . This estimation is supported by Density Functional Theory calculations, which also allow disentangling the ferro- and antiferromagnetic contributions to the exchange. Pulsed EPR experiments show that the dimer maintains relaxation times similar to the monometallic CuII porphyrins. The addressability of the two individual spins is made possible by the different g-tensors of VIV and CuII -ions, in contrast to homometallic dimers where tilting of the porphyrin planes plays a key role. Therefore, single-spin addressability in the heterometallic dimer can be maintained even with small tilting angles, as expected when deposited on surface, unlocking the full potential of molecular quantum gates for practical applications.

Probing the Structural Topology and Dynamic Properties of gp28 Using Continuous Wave Electron Paramagnetic Resonance Spectroscopy.

Lysis of Gram-negative bacteria by dsDNA phages is accomplished through either the canonical holin-endolysin pathway or the pinholin-SAR endolysin pathway. During lysis, the outer membrane (OM) is disrupted, typically by two-component spanins or unimolecular spanins. However, in the absence of spanins, phages use alternative proteins called Disruptin to disrupt the OM. The Disruptin family includes the cationic antimicrobial peptide gp28, which is found in the virulent podophage φKT. In this study, EPR spectroscopy was used to analyze the dynamics and topology of gp28 incorporated into a lipid bilayer, revealing differences in mobility, depth parameter, and membrane interaction among different segments and residues of the protein. Our results indicate that multiple points of helix 2 and helix 3 interact with the phospholipid membrane, while others are solvent-exposed, suggesting that gp28 is a surface-bound peptide. The CW-EPR power saturation data and helical wheel analysis confirmed the amphipathic-helical structure of gp28. Additionally, course-grain molecular dynamics simulations were further used to develop the structural model of the gp28 peptide associated with the lipid bilayers. Based on the data obtained in this study, we propose a structural topology model for gp28 with respect to the membrane. This work provides important insights into the structural and dynamic properties of gp28 incorporated into a lipid bilayer environment.

A nitroxide-containing cathode material for organic radical batteries studied with pulsed EPR spectroscopy

An electron spin echo in a nitroxide-containing polymer cathode film for organic radical batteries is observed for various states of charge at cryogenic temperatures. The EPR-detected state of charge (ESOC), as inferred from the number of paramagnetic centers in the film, is compared to the results of Coulomb counting based on galvanostatic charging. Spin concentration, longitudinal relaxation times T1 and phase memory times Tm strongly correlate with the ESOC. In the discharged film, the spin concentration reaches 5±3×1020 cm−3, causing a phase memory time Tm≪ 100 ns (shorter than the resonator ring-down time) that hinders the detection of the spin echo. In the charged film, the decreased spin concentration results in a longer Tm between 100 ns and 300 ns that enables spin-echo detection, yet limits the length of the microwave pulse sequence. The short, broad-band pulses cause instantaneous diffusion in the unoxidized domains across the oxidized film, affecting the relative peak intensities in the pulsed EPR spectrum. By simulating the spectral distortion caused by instantaneous diffusion, we obtain information on the local spin concentration, which complements the information on the ‘bulk’ spin concentration determined by electrochemistry and continuous-wave EPR spectroscopy.

Pulse and CW EPR Oximetry Using Oxychip in Gemcitabine-Treated Murine Pancreatic Tumors

PurposeThe goal of this work was to compare pO2 measured using both continuous wave (CW) and pulse electron paramagnetic resonance (EPR) spectroscopy. The Oxychip particle spin probe enabled longitudinal monitoring of pO2 in murine pancreatic tumor treated with gemcitabine during the course of therapy.ProceduresPancreatic PanO2 tumors were growing in the syngeneic mice, in the leg. Five doses of saline in control animals or gemcitabine were administered every 3 days, and pO2 was measured after each dose at several time points. Oxygen partial pressure was determined from the linewidth of the CW EPR signal (Bruker E540L) or from the T2 measured using the electron spin echo sequence (Jiva-25™).ResultsThe oxygen sensitivity was determined from a calibration curve as 6.1 mG/mm Hg in CW EPR and 68.5 ms−1/mm Hg in pulse EPR. A slight increase in pO2 of up to 20 mm Hg was observed after the third dose of gemcitabine compared to the control. The maximum delta pO2 during the therapy correlated with better survival.ConclusionsBoth techniques offer fast and reliable oximetry in vivo, allowing to follow the effects of pharmaceutic intervention.

Order-of-magnitude SNR improvement for high-field EPR spectrometers via 3D printed quasi-optical sample holders.

Here, we present a rapidly prototyped, cost-efficient, and 3D printed quasi-optical sample holder for improving the signal-to-noise ratio (SNR) in modern, resonator-free, and high-field electron paramagnetic resonance (HFEPR) spectrometers. Such spectrometers typically operate in induction mode: The detected EPR ("cross-polar") signal is polarized orthogonal to the incident ("co-polar") radiation. The sample holder makes use of an adjustable sample positioner that allows for optimizing the sample position to maximize the 240-gigahertz magnetic field B1 and a rooftop mirror that allows for small rotations of the microwave polarization to maximize the cross-polar signal and minimize the co-polar background. When optimally tuned, the sample holder was able to improve co-polar isolation by ≳20 decibels, which is proven beneficial for maximizing the SNR in rapid-scan, pulsed, and continuous-wave EPR experiments. In rapid-scan mode, the improved SNR enabled the recording of entire EPR spectra of a narrow-line radical in millisecond time scales, which, in turn, enabled real-time monitoring of a sample's evolving line shape.

Time-Resolved EPR Spectroscopy of Channelrhodopsin-2 Helix B Movements

The light-gated dimeric cation channel channelrhodopsin-2 (ChR2) is one of the most important optogenetic tools. Upon light activation ChR2 undergoes conformational changes, the most prominent ones include a movement of transmembrane helix B. In the present work, we apply time resolved continuous wave EPR spectroscopy to follow spectral changes of a spin label bound to position C79 located in helix B. We observed an increase of the motional freedom of the spin label side chain in illuminated ChR2. The recovery of the underlying light-induced conformational change in the dark is correlated with the recovery of the P480 state of ChR2. The observed conformational changes might be thus key elements responsible for desensitizing the channel for cation conduction.

MEPROS – Modular electron paramagnetic resonance operating software for multifunctional high-frequency EPR spectrometer

We present a software solution developed in LabVIEW for a home-built High–Frequency Electron Paramagnetic Resonance (HF-EPR) spectrometer. A modular approach was applied to control the spectrometer subsystems and simplify the adaptation to hardware changes during the development. The solution implements measuring procedures for conventional Continuous Wave EPR (CW-EPR), Frequency–Swept EPR (FS-EPR), and Two-Dimensional EPR (2D-EPR) mapping, which are relevant in different cases. The software’s automation capabilities were tested in several trial measurements to obtain CW-EPR spectra of Silicon Carbide doped by vanadium (SiC + V) at various temperatures and microwave frequencies, multi-frequency spectra via 2D-EPR mapping, and dense FS-EPR data of a lithium phthalocyanine crystal rotated in a magnetic field. Several prospective modifications of the software are discussed in the conclusion. A modular character allows the easy re-use of code portions in other experimental setups. The spectrometer and the software are currently deployed and utilized in a laboratory of EPR spectroscopy at Central European Institute of Technology (CEITEC) in Brno, and data obtained by it has been already used in a number of publications.

The impact of spin–orbit coupling on fine-structure and spin polarisation in photoexcited porphyrin triplet states

The photoexcited triplet states of porphyrins show great promise for applications in the fields of opto-electronics, photonics, molecular wires, and spintronics. The magnetic properties of porphyrin triplet states are most conveniently studied by time-resolved continuous wave and pulse electron spin resonance (ESR). This family of techniques is singularly able to probe small yet essential details of triplet states: zero-field splittings, g-anisotropy, spin polarisation, and hyperfine interactions. These characteristics are linked to spin–orbit coupling (SOC) which is known to have a strong influence on photophysical properties such as intersystem crossing rates. The present study explores SOC effects induced by the presence of Pd2+ in various porphyrin architectures. In particular, the impact of this relativistic interaction on triplet state fine-structure and spin polarisation is investigated. These properties are probed using time-resolved ESR complemented by electron-nuclear double resonance. The findings of this study could influence the future design of molecular spintronic devices. The Pd2+ ion may be incorporated into porphyrin molecular wires as a way of controlling spin polarisation.

Characterization of the active site in the thiocyanate-forming protein from Thlaspi arvense (TaTFP) using EPR spectroscopy.

Glucosinolates are plant thioglucosides, which act as chemical defenses. Upon tissue damage, their myrosinase-catalyzed hydrolysis yields aglucones that rearrange to toxic isothiocyanates. Specifier proteins such as thiocyanate-forming protein from Thlaspi arvense (TaTFP) are non-heme iron proteins, which capture the aglucone to form alternative products, e.g. nitriles or thiocyanates. To resolve the electronic state of the bound iron cofactor in TaTFP, we applied continuous wave electron paramagnetic resonance (CW EPR) spectroscopy at X-and Q-band frequencies (∼9.4 and ∼34 GHz). We found characteristic features of high spin and low spin states of a d 5 electronic configuration and local rhombic symmetry during catalysis. We monitored the oxidation states of bound iron during conversion of allylglucosinolate by myrosinase and TaTFP in presence and absence of supplemented Fe2+. Without added Fe2+, most high spin features of bound Fe3+ were preserved, while different g'-values of the low spin part indicated slight rearrangements in the coordination sphere and/or structural geometry. We also examined involvement of the redox pair Fe3+/Fe2 in samples with supplemented Fe2+. The absence of any EPR signal related to Fe3+ or Fe2+ using an iron-binding deficient TaTFP variant allowed us to conclude that recorded EPR signals originated from the bound iron cofactor.

Electron Spin Decoherence Dynamics in Magnetic Manganese Hybrid Organic-Inorganic Crystals: The Effect of Lattice Dimensionality.

Organic-inorganic metal hybrids with their tailorable lattice dimensionality and intrinsic spin-splitting properties are interesting material platforms for spintronic applications. While the spin decoherence process is extensively studied in lead- and tin-based hybrids, these systems generally show short spin decoherence lifetimes, and their correlation with the lattice framework is still not well-understood. Herein, we synthesized magnetic manganese hybrid single crystals of (4-fluorobenzylamine)2MnCl4, ((R)-3-fluoropyrrolidinium)MnCl3, and (pyrrolidinium)2MnCl4, which represent a change in lattice dimensionality from 2D and 1D to 0D, and studied their spin decoherence processes using continuous-wave electron spin resonance spectroscopy. All manganese hybrids exhibit nanosecond-scale spin decoherence time τ2 dominated by the symmetry-directed spin exchange interaction strengths of Mn2+-Mn2+ pairs, which is much longer than lead- and tin-based metal hybrids. In contrast to the similar temperature variation laws of τ2 in 2D and 0D structures, which first increase and gradually drop afterward, the 1D structure presents a monotonous rise of τ2 with the temperatures, indicating the strong correlation of spin decoherence with the lattice rigidity of the inorganic framework. This is also rationalized on the basis that the spin decoherence is governed by the competitive contributions from motional narrowing (prolonging the τ2) and electron-phonon coupling interaction (shortening the τ2), both of which are thermally activated, with the difference that the former is more pronounced in rigid crystalline lattices.

Time-resolved electron paramagnetic resonance spectrometer based on ultrawide single-sideband phase-sensitive detection.

A time-resolved electron paramagnetic resonance (TREPR) method with 40ns time resolution and a high sensitivity suitable for the detection of short-lived radicals under thermal equilibrium is developed. The key is the introduction of a new detection technique named ultrawide single sideband phase sensitive detection (U-PSD) to the conventional continuous-wave EPR, which remarkably enhanced the sensitivity for the detection of broadband transient signals compared with the direct detection protocol. By repeatedly triggering a transient kinetic event f(t) (e.g., by laser flash photolysis) under a 100kHz magnetic field modulation with precise phase control, this technique can build an ultrawide single sideband modulated signal. After single sideband demodulation, the flicker noise-suppressed signal f(t) with wide bandwidth is recovered. A U-PSD TREPR spectrometer prototype has been built, which integrated timing sequence control, laser flash excitation, data acquisition systems, and the U-PSD algorithm with a conventional continuous-wave EPR. It exhibited excellent performance in monitoring a model transient radical system, laser flash photolysis of benzophenone in isopropanol. Both the intense chemically induced dynamic electron polarization signals and the much weaker thermal equilibrium EPR signals of the generated acetone ketyl radical and benzophenone ketyl radical were clearly observed within a wide timescale ranging from sub-microsecond to milliseconds. This prototype validated the feasibility of the U-PSD technique and demonstrated its superior performance in studying complex photochemical systems containing various transient radicals, which complements the established TREPR techniques and provides a powerful tool for deep mechanistic understandings, such as in photoredox catalysis and artificial photosynthesis.

Electric-Field-Driven Spin Resonance by On-Surface Exchange Coupling to a Single-Atom Magnet.

Coherent control of individual atomic and molecular spins on surfaces has recently been demonstrated by using electron spin resonance (ESR) in a scanning tunneling microscope (STM). Here, a combined experimental and modeling study of the ESR of a single hydrogenated Ti atom that is exchange-coupled to a Fe adatom positioned 0.6-0.8nm away by means of atom manipulation is presented. Continuous wave and pulsed ESR of the Ti spin show a Rabi rate with two contributions, one from the tip and the other from the Fe, whose spin interactions with Ti are modulated by the radio-frequency electric field. The Fe contribution is comparable to the tip, as revealed by its dominance when the tip is retracted, and tunable using a vector magnetic field. The new ESR scheme allows on-surface individual spins to be addressed and coherently controlled without the need for magnetic interaction with a tip. This study establishes a feasible implementation of spin-based multi-qubit systems on surfaces.

Towards high performance dielectric microwave resonators for X-band EPR spectroscopy

Microwave (MW) resonators in Electron Paramagnetic Resonance (EPR) spectroscopy concentrate the MW magnetic field (B1) at the sample and separate the MW electric field from the sample. There are numerous experimental methods in EPR spectroscopy which all impose different requirements on MW resonators (e.g. high or low quality factor, MW conversion, and B1-field homogeneity). Although commercial spectrometers offer standardized MW resonators for a broad application range, newly emerging and highly-specialized research fields push these spectrometers to or beyond their sensitivity limits. Optimizing the MW resonator offers one direct approach to improve the sensitivity. Here we present three low-cost optimization approaches for a commercially available X-band (9–10 GHz) MW resonator for three experimental purposes (continuous-wave (CW), transient and pulse EPR). We obtain enhanced MW conversion factors for all three optimized resonators and higher quality factors for two optimized resonators. The latter is important for CW and transient EPR. Furthermore, we fabricated a resonator which features an extended area of homogeneous B1-field and, hence, improved pulse EPR performance. Our results demonstrate that small changes to a commercial MW resonator can enhance its performance in general or for specific applications.

Anisotropic Multiexciton Quintet and Triplet Dynamics in Singlet Fission via Pulsed Electron Spin Resonance.

The quintet triplet-pair state may be generated upon singlet fission and is a critical intermediate that dictates the fate of excitons, which can be exploited for photovoltaics, information technologies, and biomedical imaging. In this report, we demonstrate that continuous-wave and pulsed electron spin resonance techniques such as phase-inverted echo-amplitude detected nutation (PEANUT), which have emerged as the primary tool for identifying the spin pathways in singlet fission, probe fundamentally different triplet-pair species. We directly observe that the generation rate of high-spin triplet pairs is dependent on the molecular orientation with respect to the static magnetic field. Moreover, we demonstrate that this observation can prevent incorrect analysis of continuous-wave electron spin resonance (cw-ESR) measurements and provide insight into the design of materials to target specific pathways that optimize exciton properties for specific applications.

Continuous-wave and pulsed EPR studies of glass-like carbon with high spin concentration: Evidence for triplet states

Glass-like carbons (GLCs) are a large group of disordered carbon materials which have applications in various fields, but not all their properties are well characterized. In particular, they have high concentration of paramagnetic centers (PCs) which are easily observed by electron paramagnetic resonance (EPR). However, an apparently simple EPR spectrum, together with its peculiarities caused by high electrical conductivity, make it difficult to identify the PCs and their origin remains unknown. To resolve these unsettled questions, in this work we use the vacuum Rabi splitting and Rabi oscillations which so far have not been applied to study these materials by the continuous wave (CW) and pulsed EPR. GLC produced by pyrolysis of phenol-formaldehyde resin at 1120 K was studied in the temperature range of 4.3–300 K. Using the Rabi splitting we found conditions for undistorted registration of CW EPR spectra and determined spin concentration (1×1020 spin/g). The temperature dependence of the EPR susceptibility shows the coexistence of Pauli paramagnetism due to conduction electrons and the Curie-type paramagnetism of thermally activated PCs with the activation energy of 5.7 cm-1. These PCs may be excited triplet states, the presence of which is revealed by Rabi oscillations in the pulsed EPR.

Conformational changes in the activation loop of a bacterial PASTA kinase.

Many bacterial genomes encode a transmembrane protein kinase belonging to the PASTA kinase family, which controls numerous processes in diverse bacterial pathogens, including antibiotic resistance, cell division, stress resistance, toxin production, and virulence. PASTA kinases share a conserved three-part domain architecture, consisting of an extracellular PASTA domain, proposed to sense the peptidoglycan layer status, a single transmembrane helix, and an intracellular Ser/Thr kinase domain. The crystal structures of the kinase domain from two homologous PASTA kinases reveal a characteristic two-lobed structure typical of eukaryotic protein kinases with a centrally located, but unresolved, activation loop that becomes phosphorylated and regulates downstream signaling pathways. We previously identified three sites of phosphorylation on the activation loop (T163, T166, and T168) of IreK, a PASTA kinase from the pathogen Enterococcus faecalis, as well as a distal phosphorylation site (T218) that each influence IreK activity in vivo. Still, the mechanism by which loop phosphorylation regulates PASTA kinase function is yet unknown. Therefore, we utilized site-directed spin labeling (SDSL) and continuous wave (CW) electron paramagnetic resonance (EPR) spectroscopy to assess the E. faecalis IreK kinase activation loop dynamics, including the effects of phosphorylation on activation loop motion, and the IreK-IreB interaction. Our results reveal that the IreK activation loop occupies a more immobile state when dephosphorylated, and that loop autophosphorylation shifts the loop to a more mobile state that can then enable interaction with IreB, a known substrate.

Refining the Dynamic Network of T2SS Endopilus Tip Heterocomplex Combining cw-EPR and Nitroxide-GdIII Distance Measurements.

The type 2 secretion system (T2SS) is a bacterial nanomachine composed of an inner membrane assembly platform, an outer membrane pore and a dynamic endopilus. T2SS endopili are organized into a homo-multimeric body formed by the major pilin capped by a heterocomplex of four minor pilins. The first model of the T2SS endopilus was recently released, even if structural dynamics insights are still required to decipher the role of each protein in the full tetrameric complex. Here, we applied continuous-wave and pulse EPR spectroscopy using nitroxide-gadolinium orthogonal labelling strategies to investigate the hetero-oligomeric assembly of the minor pilins. Overall, our data are in line with the endopilus model even if they evidenced conformational flexibility and alternative orientations at local scale of specific regions of minor pilins. The integration of different labelling strategies and EPR experiments demonstrates the pertinence of this approach to investigate protein-protein interactions in such multiprotein heterocomplexes.

Generation and Characterization of a Tetraradical Embedded in a Curved Cyclic Paraphenylene Unit.

Unique spin-spin (magnetic) interactions, ring-size effects on ground-state spin multiplicity, and in-plane aromaticity has been found in localized 1,3-diradicals embedded in curved benzene structures such as cycloparaphenylene (CPP). In this study, we characterized the magnetic interactions in a tetraradical consisting of two localized 1,3-diradical units connected by p-quaterphenyl within a curved CPP skeleton by electron paramagnetic resonance (EPR) spectroscopy and quantum chemical calculations. Persistent triplet species with zero-field splitting parameters similar to those of a triplet 1,3-diphenylcyclopentane-1,3-diyl diradical were observed by continuous wave (CW) or pulsed X-band EPR measurements. The quintet state derived from the ferromagnetic interaction between the two triplet diradical moieties was not detected at 20 K under glassy matrix conditions. At the B3LYP/6-31G(d) level of theory, the singlet state was lower in energy than the triplet and quintet states. These findings will aid in the development of open-shell species for material science application.