HYSCORE and DEER with an upgraded 95GHz pulse EPR spectrometer

The set-up of a new microwave bridge for a 95 GHz pulse EPR spectrometer is described. The virtues of the bridge are its simple and flexible design and its relatively high output power (0.7 W) that generates pi pulses of 25 ns and a microwave field, B(1)=0.71 mT. Such a high B(1) enhances considerably the sensitivity of high field double electron-electron resonance (DEER) measurements for distance determination, as we demonstrate on a nitroxide biradical with an interspin distance of 3.6 nm. Moreover, it allowed us to carry out HYSCORE (hyperfine sublevel-correlation) experiments at 95 GHz, observing nuclear modulation frequencies of 14N and 17O as high as 40 MHz. This opens a new window for the observation of relatively large hyperfine couplings, yet not resolved in the EPR spectrum, that are difficult to observe with HYSCORE carried out at conventional X-band frequencies. The correlations provided by the HYSCORE spectra are most important for signal assignment, and the improved resolution due to the two dimensional character of the experiment provides 14N quadrupolar splittings.     

Pulsed 95 GHz high-field EPR heterodyne spectrometer with high spectral and time resolution

Various time resolved EPR methods are applied to different test samples to demonstrate the abilities of pulsed high-field EPR spectroscopy. Two-pulse-echo field swept EPR spectroscopy on a nitroxide radical shows the increased spectral resolution by separating different spin systems by their relaxation properties. Additionally N¹⁴ electron-spin-echo-envelope-modulation (ESEEM) is observed for these systems at fields as high as 3.5 T. Thus, the N¹⁴ hyperfine interaction couplings can be probed by ESEEM and pulsed ENDOR (electron-nuclear-double-resonance) experiments. The sensitivity of pulsed ENDOR experiments is compared with cw-ENDOR. The different linewidths and amplitudes of the two methods are discussed. Transient nutation experiments on light induced triplet states demonstrate the high sensitivity and time resolution of high-field pulsed EPR. The sensitivity and time resolution of our 95 GHz spectrometer are determined and compared with pulsed X-band EPR spectrometer performances.     

Fourier-transform EPR at high-field/high-frequency (3.4 T/95 GHz) using broadband stochastic microwave excitation

Stochastic excitation with a full-width-half-maximum bandwidth of 250 MHz was used to perform Fourier-transform (FT) high-field/high-frequency electron paramagnetic resonance (EPR) at 3.4T/95 GHz (W-band). Thereby, the required microwave peak power is reduced by a factor of tau(p)/T1 as compared to equivalent pulsed FT EPR in which the spin system with spin-lattice relaxation time T1 is excited by a single microwave pulse of length tau(p). Stochastic EPR is particularly interesting under high-field/high-frequency conditions, because the limited output power of mm microwave sources, amplifiers, and mixers makes pulse FT EPR in that frequency domain impossible, at least for the near future. On the other hand, FT spectroscopy offers several advantages compared to field-swept magnetic resonance methods, as is demonstrated by its success in NMR and X-band EPR. In this paper we describe a novel stochastic W-band microwave bridge including a bimodal induction mode transmission resonator that serves for decoupling the microwave excitation and signal detection. We report first EPR measurements and discuss experimental difficulties as well as achieved sensitivity. Moreover, we discuss future improvements and the possibility for an application of stochastic W-band FT EPR to transient signals such as those of photoexcited radical pairs in photosynthetic reaction centers.     

High-field/high-frequency EPR spectrometer operating in pulsed and continuous-wave mode at 180 GHz

A novel electron paramagnetic resonance (EPR) spectrometer is reported, which has been developed to allow pulsed EPR experiments with high sensitivity and time resolution at a microwave (MW) frequency of 180 GHz (G-band) and wavelengths of approximately 1.6 mm. This corresponds to a magnetic field of about 6.4 T forg ≈ 2 signals. The “hybrid” system architecture combines components of quasioptical as well as conventional MW techniques, making it possible to achieve excellent spectrometer performance with respect to sensitivity and time resolution. Quasioptical MW components have been used to design an MW circulator allowing high sensitivity and low bias operation in the reflection mode. A miniaturized, closed-type cylindrical cavity provides a high sample filling factor and an adequate MW field strength (B₁) enhancement and thus permits reasonably short MW pulses (60 ns for a π/2 pulse) even with a moderate MW input power (15 mW at the cavity). Commercial quartz capillaries (up to 0.5 mm internal diameter) can be used as sample holders for a broad range of applications.     

Microwave Induced Optical Nuclear Polarization at 75 GHz: A quantitative analysis

A quantitative analysis of Microwave Induced Optical Nuclear Polarization (MIONP) on crystals of fluorene-h₁₀ doped with phenanthrene-d₁₀ at 75 GHz and 1.4 K is presented. Two effects are studied in detail: the nuclear spin diffusion barrier and the phonon bottleneck. Experiments are presented that allow the identification of the proton spins on fluorene-h₁₀ molecules located inside the nuclear spin diffusion barrier surrounding phenanthrene-d₁₀ molecules excited in the photo-excited triplet state. Using this result the experimental values for the triplet spin-lattice relaxation (SLR) rates and the nuclear SLR rate can be related to each other without any fitting parameter. Microwave frequency and magnetic field modulation are used during MIONP to prove that the triplet SLR is phonon-bottlenecked. Subsequently a quantitative analysis of MIONP in the system fluorene-h₁₀ doped with phenanthrene-d₁₀ is obtained.     

Low-Field, Time-Resolved Dynamic Nuclear Polarization with Field Cycling and High-Resolution NMR Detection

Dynamic nuclear polarization (DNP) effects in aqueous solutions of stable 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) radicals were studied in a pulsed mode of pumping the electron paramagnetic resonance (EPR) transitions. Our fast field cycling setup allowed us to perform the EPR pumping at low magnetic fields and to detect the enhanced nuclear magnetic resonance signals at 7 T with high spectral resolution. Pumping was performed at two different frequencies, 300 MHz and 1.4 GHz, corresponding to magnetic fields around 10 and 48.6 mT, respectively. For both frequencies, the DNP enhancements were close to the limiting theoretical values of ?110 (¹⁴N TEMPOL) and ?165 (¹⁵N TEMPOL). Our pulsed experiments exploit coherent motion of the electronic spins in the radio-frequency field as seen by an oscillatory component in the dependence of the DNP effect on the radio-frequency pulse duration. The DNP enhancement was studied in detail as a function of the pulse length, duty cycle, delay between the pulses, and applied power. The analysis of the results shows that pulsed DNP experiments provide an opportunity to achieve enhancements of about ?110 with relatively low applied power as compared to the standard continuous-wave DNP experiments. An adequate theoretical approach to the problem under study is suggested.     

脉冲动态核极化增强的NMR 和MRI 系统研究

该文介绍了一种自行设计和构建的可扩展脉冲动态核极化谱仪,可以实现核磁共振波谱与磁共振成像的功能．该仪器的新颖设计主要有：1) 采用基于PCIe 的分布式总线结构,能够极大地提高数据传输效率和通信可靠性,实现精确控制脉冲序列;2) 采用外部高速的DDR 芯片存储脉冲序列元素和FID 数据,可以极大的提高脉冲序列的执行速度,减少快速成像序列的TR 时间间隔;3) 采用时钟移相技术,可以精确产生分辨率为纳秒级别的数字脉冲．最后对该仪器的动态核极化-磁共振波谱与核磁共振成像功能进行了实验验证．     

A spectrometer for EPR,DNP, and multinuclear high-resolution NMR

We describe a magnetic resonance spectrometer capable of EPR, dynamic nuclear polarization, and multinuclear high-resolution NMR. The operating field is 1.4 T, corresponding to Larmor frequencies of 40 GHz and 60 MHz for electrons and protons, respectively. The microwave side of the probe is based on a Fabry-Perot resonator (FPR ), an open structure that enhances power-to-field conversion for efficient saturation of the EPR for dynamic polarization, and further permits in situ detection for EPR. This allows the external field to be set at, rather than scanned for, the optimal DNP position. Moreover, we have found that adjustments necessary for maximizing DNP may be done via optimization of the EPR signal, a feature of particular significance for samples which exhibit NMR signals on the borderline of detectability, i.e., samples for which DNP is of special importance. 'H and '3C polarization enhancements achieved using the FPR are compared with devices used by others, in particular the horn /reflector system used by Wind and co-workers. Direct '3C enhancements large enough to detect 2.5 x 10'6 spins in (fluoranthenyl)2 PF6 after a single one-second polarization period have been obtained, and the first high-field 'Li DNP results are also presented.     

Direct EPR detection of transient and continuous wave signals at 2.5 GHz

Direct detection of free induction decays and electron spin echoes, and the recording of echo-detected EPR spectra and electron spin echo envelope modulation patterns at a microwave frequency of 2.5 GHz is demonstrated. This corresponds to the measurement of the transverse magnetization in the laboratory frame, rather than in the rotating frame as usually done by down-converting the signal (homodyne detection). An oscilloscope with a 6-GHz analog bandwidth, a sampling rate of 20 GigaSamples per second, and a trigger frequency of 5 GHz for the edge trigger and 750 MHz for the advanced trigger, is used in these experiments. For signal averaging a 3-GHz microwave clock divider has been developed to synchronize the oscilloscope with the frequency of the EPR signal. Moreover, direct detection of continuous wave EPR signals at 2.5 GHz is described.     

A CW and pulse EPR spectrometer operating at 122 and 244 GHz using a quasi-optical bridge and a cryogen-free 12 T superconducting magnet

A high-field continuous-wave (CW) and pulse electron paramagnetic resonance spectrometer operating at 122 and 244 GHz is described. The instrument is based on a millimeter-wave bridge built from quasi-optical components. To improve the sensitivity, a cryo-cooled detector/mixer is used. The magnetic field is generated using a cryogen-free superconducting 12 T magnet (warm bore, 88 mm) equipped with a helium-flow cryostat for sample cooling. The advantages of this spectrometer are described and first results (obtained in CW mode) on different types of samples at 122 and 244 GHz are presented. The extensions to pulse operation as well as double resonance techniques (electron-electron and electron-nuclear) are briefly discussed.     

A continuous-wave and pulsed electron spin resonance spectrometer operating at 275 GHz

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.     

TM110模式DNP探头的研制与应用

为了满足自主研发的X波段动态核极化系统(DNP-NMR/MRI)对探头的需求,设计制作了用于动态核极化的TM_(110)模式圆柱形谐振腔探头.通过理论分析计算得到了探头腔体的初步尺寸,进而利用软件对其完整结构做进一步的仿真优化,并测试了基于优化参数设计加工的探头的性能参数.完成了探头在自主研制的场强为0.35 T的DNP-NMR/MRI系统上的测试,得到了大于50倍信噪比(S/N)增强的质子信号,同时获得信噪比增强的磁共振影像.     

Fast Dissolution Dynamic Nuclear Polarization NMR of 13C-Enriched 89Y-DOTA Complex: Experimental and Theoretical Considerations

The yttrium complex of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(1′-¹³C-acetic acid) [¹³C]DOTA was synthesized. Fast dissolution dynamic nuclear polarization (DNP) nuclear magnetic resonance (NMR) studies revealed that the ⁸⁹Y, ¹³C, and ¹⁵N nuclei present in the complex could be co-polarized at the same optimum microwave irradiation frequency. The liquid-state spin–lattice relaxation time T ₁ of these nuclei were found to be reasonably long to preserve some or most of the DNP-enhanced polarization after dissolution. The hyperpolarized ¹³C and ⁸⁹Y NMR signals were optimized in different glassing mixtures. The overall results are discussed in light of the thermal mixing model of DNP.     

A Q-band pulse EPR/ENDOR spectrometer and the implementation of advanced one- and two-dimensional pulse EPR methodology

A versatile high-power pulse Q-band EPR spectrometer operating at 34.5--35.5 GHz and in a temperature range of 4--300 K is described. The spectrometer allows one to perform one- and two-dimensional multifrequency pulse EPR and pulse ENDOR experiments, as well as continuous wave experiments. It is equipped with two microwave sources and four microwave channels to generate pulse sequences with different amplitudes, phases, and carrier frequencies. A microwave pulse power of up to 100 W is available. Two channels form radiofrequency pulses with adjustable phases for ENDOR experiments. The spectrometer performance is demonstrated by single crystal pulse ENDOR experiments on a copper complex. A HYSCORE experiment demonstrates that the advantages of high-field EPR and correlation spectroscopy can be combined and exploited at Q-band. Furthermore, we illustrate how this combination can be used in cases where the HYSCORE experiment is no longer effective at 35 GHz because of the shallow modulation depth. Even in cases where the echo modulation is virtually absent in the HYSCORE experiment at Q-band, matched microwave pulses allow one to get HYSCORE spectra with a signal-to-noise ratio as good as at X-band. Finally, it is shown that the high microwave power, the short pulses, and the broad resonator bandwidth make the spectrometer well suited to Fourier transform EPR experiments.     

Solid-Effect Rate for Dynamic Nuclear Polarization of Spin-l/2 and Spin-1 Nuclei

The solid-effect rate equations for the dynamic nuclear polarization of spin-1/2 (tritium or hydrogen) or spin-1 (deuterium) nuclei are derived in the limiting case of the electronspin-resonance line width being much smaller than the nuclear-magnetic-resonance frequency. The exact analytical solutions of the rate equations for spin-1/2 nuclei are given, while the rate equations for spin-1 nuclei, which are nonlinear, are solved in decoupling approximation. The nuclear polarization and the electronic polarization as well as the times to polarization are calculated in low temperature limit. It is found that there is an optimal radio frequency pumping rate for the nuclear polarizations, which is not considered until now.     

Investigating magnetically aligned phospholipid bilayers with EPR spectroscopy at Q-band (35 GHz): optimization and comparison with X-band (9 GHz)

This paper presents the improvement and advantages of investigating magnetically aligned phospholipid bilayers (bicelles) utilizing electron paramagnetic resonance (EPR) spectroscopy at a microwave frequency of 35 GHz (Q-band) and at a high magnetic field strength of 1.25 T when compared to weaker magnetic fields for X-band EPR studies. The nitroxide spin label 3beta-doxyl-5alpha-cholestane (cholestane or CLS) was inserted into the bicelles and utilized to demonstrate the effects of macroscopic bilayer alignment through the measurement of orientational dependent hyperfine splittings. The effects of different lanthanide ions with varying degree of magnetic susceptibility anisotropy were examined. The requirement of minimal amounts of the Tm3+ and Dy3+ lanthanide ions for well-aligned bicelles were examined for Q-band and compared with amounts required for X-band bicelle alignment studies. At a magnetic field of 1.25 T (when compared to 0.63 T at X-band), the perpendicular and parallel orientation were aligned with lower concentrations of Dy3+ and Tm3+, respectively, and thereby eliminating/minimizing the unwanted effects associated with lanthanide-protein interactions. Thus, it is much easier to magnetically align phospholipid bilayers at Q-band when compared to X-band.