Time-resolved electron paramagnetic resonance of radical pair intermediates in cryptochromes

Electron transfer plays a key role in many biological systems, including core complexes of photosynthesis and respiration. As this involves unpaired electron spins, electron paramagnetic resonance (EPR) is the method of choice to investigate such processes. Systems that show photo-induced charge separation and electron transfer are of particular interest, as here the processes can easily be synchronised to the experiment and therefore followed directly over its time course. One particular class of proteins, the cryptochromes, showing charge separation and in turn spin-correlated radical pairs upon excitation with blue light, have been investigated by time-resolved EPR spectroscopy in great detail and the results obtained so far are summarised in this contribution. Highlights include the first observation of spin-correlated radical pairs in these proteins, a fact with great impact on the proposed role as key part of a magnetic compass of migratory birds, as well as the assignment of the radical-pair partners and the unravelling of alternative and unexpected electron transfer pathways in these proteins, giving new insights into aspects of biological electron transfer itself.     

Carbon-based standards for electron paramagnetic resonance spectroscopy

In order to meet the need for a good new EPR intensity andg-value standard whose paramagnetic species are carbon-based radicals, several materials were investigated, including coal, fusinite (a coal maceral), and several carbohydrate chars. Of the prototypical standards prepared, a chemically-treated fusinite is recommended as most suitable because of its chemical stability, spin density, EPR signal line shape and line width, microwave power saturation characteristics, availability, and homogeneity. Effects of dilution with KBr, KCl, and polymer are negligible, although the line width is broadened in the presence of paramagnetic gases. Several model standard compounds have been prepared in a polymer matrix to minimize changes in packing density over time.     

Tunable-frequency high-field electron paramagnetic resonance

A tunable-frequency methodology based on backward wave oscillator sources in high-frequency and -field EPR (HFEPR) is described. This methodology is illustrated by an application to three non-Kramers transition metal ion complexes and one Kramers ion complex. The complexes are of: Ni(II) (S=1) as found in dichlorobistriphenylphosphanenickel(II), Mn(III) (S=2) as found in mesotetrasulfonatoporphyrinatomanganese(III) chloride, Fe(II) (S=2) as found in ferrous sulfate tetrahydrate, and Co(II) (S=3/2) as found in azido(tris(3-tert-butylpyrazol-1-yl)hydroborate)cobalt(II). The above Ni(II) and Mn(III) complexes have been studied before by HFEPR using the multifrequency methodology based on Gunn oscillator sources, but not by the present method, while the Fe(II) and Co(II) complexes presented here have not been studied by any form of HFEPR. Highly accurate spin Hamiltonian parameters can be obtained by the experimental methodology described here, in combination with automated fitting procedures. This method is particularly successful in determining g-matrix parameters, which are very difficult to extract for high-spin systems from single frequency (or a very limited set of multi-frequency) HFEPR spectra, but is also able to deliver equally accurate values of the zero-field splitting tensor. The experimental methods involve either conventional magnetic field modulation or an optical modulation of the sub-THz wave beam. The relative merits of these and other experimental methods are discussed.     

Transmission electron spin resonance as a probe of the metallic interface

Observation of coupled TESR in bilayers of dissimilar pure metals is reported. Positive entropy production and detailed balance are incorporated in general linear relations between non-equilibrium spin densities and currents at the interface. Spin transport coefficients for samples exhibiting both strong and weak coupling are deduced from the data by comparison with computer generated lineshapes.     

Synthesis and electron paramagnetic resonance study of a nitroxide free radical covalently bonded on aminopropyl-silica gel

A solid spin-labeled material was obtained starting from 2-chloro-3,5-dinitro-N-(4-(2,2,6,6-tetramethyl-piperidine-1-oxyl)-benzamide) and aminopropyl-silica gel. Stability tests showed that even after several months the spin-labeled material had the same properties as immediately after synthesis. EPR properties of the TEMPO-derivatized silica were studied as a function of solvent polarity and temperature. Rotational correlation times were calculated from EPR spectra and correlated with solvent characteristics and temperature. Polar solvents induce a fast motion of the spin-label, clearly seen in the EPR spectra by the apparition of the well-known TEMPO radical triplet. The solid spin-labeled (dry) sample showed a high interspin interaction, which can be disrupted not only by different (liquid) solvents, but also by absorption of different solids, like cyclodextrins, dendrimers or polyethyleneglycols. Also, changes induced by the temperature were studied in the case of toluene wet sample. From 150 to 370 K, the spectrum is changing from a slow motion spectrum type to a fast motion regime. The preparative procedures to obtain the spin-labeled silica as well as some of its parameters are described.     

Nuclear polarization by electron paramagnetic resonance in paramagnetic crystals

The possibility of the polarization of nuclei in paramagnetic salts by saturation of electron paramagnetic resonance is theoretically analyzed. The proposed method assumes saturation of the forbidden transition of the typeM=±1, m=±1, ±2, for mutually perpendicular external magnetic and high-frequency fields. The analysis is carried out for the case of a large quadrupole moment of the nucleus. The degree of orientation attained is comparable in order of magnitude with Overhauser's method. This method is particularly suitable for the polarization of nuclei of transuranium elements.

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. M=± 1, m==±1, ±2 . . . .

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In conclusion the author would like to thank J. Burget, J. ajko, M. Kolá and M. ott for helping in the laborious solution of system (16).     

Low-power electron paramagnetic resonance spin-echo spectroscopy

Electron spin-echo experiments generally require microwave power levels of hundreds of watts to produce the 5–10 G of RF field to generate 90° and 180° pulses in 10 ns. A low-power (i.e., less than I W) EPR spectrometer using a loop-gap resonator can generate the full range of time-domain experiments on samples with submicrosecond recovery times; 90° pulses are generated in 40 ns, and relaxation times as short as 22 ns are measured. Appropriate time-domain experiments were performed to independently measure the spinspin relaxation time, phase memory time, and spin-lattice relaxation time; the results were compared with CW saturation. It was found that the spin-spin and spin-lattice relaxation rates do differ by about 5%. The entire CW signal of PADS is reconstructed from a pulse experiment at a single field position. Small differences in linewidths among the three lines were seen in accordance with theory.     

Cryostat for low temperature x-irradiation and electron paramagnetic resonance

A cryostat for x-irradiation of solid samples at liquid air temperature is described. The design of the sample mount in the cryostat enables quick transfer of the sample after x-irradiation into EPR cavity to facilitate EPR study without warming the sample.     

Data acquisition card for varian electron paramagnetic resonance spectrometers

An easy to assemble and inexpensive PC card that interfaces E-Line Varian spectrometers (E-6, E-9, E-12 and E-15) with IBM compatible PCs is presented. The card has a Programmable Interval Timer so it does not need an external clock making it instrument independent. The accessory contains its own RAM. This allows the device to store the spectrum while sampling is in progress, so the PC can meanwhile be used for any other task.     

Multifrequency electron paramagnetic resonance of ultramarine blue

Electron paramagnetic resonance spectra of the S^3/? radical center in ultramarine blue over a factor of about 2500 in frequency (258 MHz to 670 GHz) reveal a substantially Lorentzian shape, without resolution of g anisotropy. Variable temperature measurements found that the line width is independent of temperature, within experimental uncertainty, up to about 90 K at 9.5 GHz and between ca. 5 K and room temperature at 95 and 217 GHz, as expected for an exchange-narrowed signal. Analysis of the increase in the low-temperature line width as a function of frequency above 9 GHz is consistent with an exchange interaction of about 2· 10^?2 K. The line width increases as frequency is decreased from 2.7 GHz to 258 MHz which is attributed to the contribution from nonsecular terms that has been denoted the “10/3” effect.     

Precisiong-factor measurements of electron paramagnetic resonance standards

An attempt is made to measureg-factors of electrons in EPR standard samples with the lowest obtainable uncertainty. An experimental equipment for preciseg-factor measurements is described, working at microwave frequencies in the 9 and 36 GHz region, and at room temperature. The course of the measuring process is mentioned in some detail and the sources of errors, which arise during the measurement, are discussed. As an example, the determination ofg-factors of certain EPR standards (using DPPH single crystals and powdered charred dextrose) is presented. No dispersion ofg-factor value has been observed between 9 and 36 GHz.     

Targeted-ROI imaging in electron paramagnetic resonance imaging

Electron paramagnetic resonance imaging (EPRI) is a technique that has been used for in vivo oxygen imaging of small animals. In continuous wave (CW) EPRI, the measurement can be interpreted as a sampled 4D Radon transform of the image function. The conventional filtered-backprojection (FBP) algorithm has been used widely for reconstructing images from full knowledge of the Radon transform acquired in CW EPRI. In practical applications of CW EPRI, one often is interested in information only in a region of interest (ROI) within the imaged subject. It is desirable to accurately reconstruct an ROI image only from partial knowledge of the Radon transform because acquisition of the partial data set can lead to considerable reduction of imaging time. The conventional FBP algorithm cannot, however, reconstruct accurate ROI images from partial knowledge of the Radon transform of even dimension. In this work, we describe two new algorithms, which are referred to as the backprojection filtration (BPF) and minimum-data filtered-backprojection (MDFBP) algorithms, for accurate ROI-image reconstruction from a partial Radon transform (or, truncated Radon transform) in CW EPRI. We have also performed numerical studies in the context of ROI-image reconstruction of a synthetic 2D image with density similar to that found in a small animal EPRI. This demonstrates both the inadequacy of the conventional FBP algorithm and the success of BPF and MDFBP algorithms in ROI reconstruction. The proposed ROI imaging approach promises a means to substantially reduce image acquisition time in CW EPRI.     

Lattice dynamic effects in electron paramagnetic resonance

The electron-phonon interaction for a paramagnetic impurity in an insulating crystal is derived and from that an electron-electron interaction in a higher order is considered. An attempt is made to provide a unified presentation of the effect of phonons on the parameters measured in an electron-paramagnetic resonance experiment. The phonon-induced corrections to the zero-field splitting, the hyperfine field, the superhyperfine splitting and line intensities is reviewed theoretically along with experimental data. Both the point charge and covalent models are discussed. The effect of local and resonance modes is calculated and discussed in terms of isotope effect. Some related problems of the energy levels, dispersion, screening and phonon force are also included in appendices.     

Observation of helicon-excited electron paramagnetic resonance in a high-mobility semiconductor

Helicon waves can be exploited in EPR studies of localized magnetic moments in high-conductivity semiconductors when the free carriers have small effective mass and high mobility. In an electron plasma helicons have precisely the polarization required to elicit EPR, and the plasma background actually enhances the resonance intensity. Microwave transmission experiments on Hg_1-xMn_xTe, a narrow-gap semiconductor with localized Mn⁺⁺ magnetic moments, are described to illustrate this effect.     

Conduction electron spin resonance as a probe of zinc diffusion in lithium films

CESR is used to measure the maximum solubility C₀ and the diffusion constant D of zinc in lithium. The sample is a bimetallic layer obtained by thermal evaporation. The first deposited metal (zinc) is 300 Å thick, the second (lithium) 2μm thick. We analyze the resonance lineshape, taking into account the zinc diffusion via the Fick theorem. From the variations of the Li resonance linewidth as a function of time, we measure C₀ = 50 ppm and D = 160 Ų sec^-1 at room temperature.     

An X-band time domain electron paramagnetic resonance spectrometer

A computer-controlled X-band time domain electron paramagnetic resonance (EPR) spectrometer, with a time resolution of the order of 0.5μsec, has been constructed with many of the crucial microwave components designed and fabricated by the Microwave Engineering Group of TIFR. The spectrometer operates either in a microwave power pulsed mode for determination of spin-lattice relaxation times by the saturation recovery technique or in the kinetic mode for determination of the time dependence of EPR signal after laser excitation. It has an automatic frequency control, an automatic phase control and, most importantly, a field-frequency lock which ensures good stability of the EPR line positions enabling signal averaging for extended periods. The constructional details of the spectrometer and its performance in both the modes are described here by reporting results on certain typical systems.