A tunable reentrant resonator with transverse orientation of electric field for in vivo EPR spectroscopy

There has been a need for development of microwave resonator designs optimized to provide high sensitivity and high stability for EPR spectroscopy and imaging measurements of in vivo systems. The design and construction of a novel reentrant resonator with transversely oriented electric field (TERR) and rectangular sample opening cross section for EPR spectroscopy and imaging of in vivo biological samples, such as the whole body of mice and rats, is described. This design with its transversely oriented capacitive element enables wide and simple setting of the center frequency by trimming the dimensions of the capacitive plate over the range 100-900 MHz with unloaded Q values of approximately 1100 at 750 MHz, while the mechanical adjustment mechanism allows smooth continuous frequency tuning in the range +/-50 MHz. This orientation of the capacitive element limits the electric field based loss of resonator Q observed with large lossy samples, and it facilitates the use of capacitive coupling. Both microwave performance data and EPR measurements of aqueous samples demonstrate high sensitivity and stability of the design, which make it well suited for in vivo applications.     

Development of a resonator with automatic tuning and coupling capability to minimize sample motion noise for in vivo EPR spectroscopy

EPR spectroscopy has been applied to measure free radicals in vivo; however, respiratory, cardiac, and other movements of living animals are a major source of noise and spectral distortion. Sample motions result in changes in resonator frequency, Q, and coupling. These instabilities limit the applications that can be performed and the quality of data that can be obtained. Therefore, it is of great importance to develop resonators with automatic tuning and automatic coupling capability. We report the development of automatic tuning and automatic coupling provisions for a 750-MHz transversely oriented electric field reentrant resonator using two electronically tunable high Q hyperabrupt varactor diodes and feedback loops. In both moving phantoms and living mice, these automatic coupling control and automatic tuning control provisions resulted in an 8- to 10-fold increase in signal-to-noise ratio.     

Detection of electron paramagnetic resonance absorption using frequency modulation

A frequency modulation (FM) method was developed to measure electron paramagnetic resonance (EPR) absorption. The first-derivative spectrum of 1,1-diphenyl-2-picrylhydrazyl (DPPH) powder was measured with this FM method. Frequency modulation of up to 1.6 MHz (peak-to-peak) was achieved at a microwave carrier frequency of 1.1 GHz. This corresponds to a magnetic field modulation of 57microT (peak-to-peak) at 40.3 mT. By using a tunable microwave resonator and automatic control systems, we achieved a practical continuous-wave (CW) EPR spectrometer that incorporates the FM method. In the present experiments, the EPR signal intensity was proportional to the magnitude of frequency modulation. The background signal at the modulation frequency (1 kHz) for EPR detection was also proportional to the magnitude of frequency modulation. An automatic matching control (AMC) system reduced the amplitude of noise in microwave detection and improved the baseline stability. Distortion of the spectral lineshape was seen when the spectrometer settings were not appropriate, e.g., with a lack of the open-loop gain in automatic tuning control (ATC). FM is an alternative to field modulation when the side-effect of field modulation is detrimental for EPR detection. The present spectroscopic technique based on the FM scheme is useful for measuring the first derivative with respect to the microwave frequency in investigations of electron-spin-related phenomena.     

1.1-GHz continuous-wave EPR spectroscopy with a frequency modulation method

Continuous-wave EPR spectroscopy using a frequency modulation (FM) scheme was developed. An electronically tunable resonator and an automatic tuning control (ATC) system were used. Using the FM scheme instead of magnetic field modulation, we detected EPR absorption at the first derivative mode. We used a microwave frequency of 1.1 GHz in the present experiment. Similar signal-to-noise ratios were obtained with conventional field modulation and the FM method, and a low-quality factor EPR resonator was not necessary to suppress the significant microwave reflection from the resonator. The FM method with a tunable resonator may be an alternative solution to achieving phase-sensitive detection, when the side-effects of magnetic field modulation, such as microphonic noise and mechanical vibration, are detrimental for EPR detection.     

Development of a Resonator with Automatic Tuning and Coupling Capability to Minimize Sample Motion Noise for in Vivo EPR Spectroscopy

EPR spectroscopy has been applied to measure free radicals in vivo; however, respiratory, cardiac, and other movements of living animals are a major source of noise and spectral distortion. Sample motions result in changes in resonator frequency, Q, and coupling. These instabilities limit the applications that can be performed and the quality of data that can be obtained. Therefore, it is of great importance to develop resonators with automatic tuning and automatic coupling capability. We report the development of automatic tuning and automatic coupling provisions for a 750-MHz transversely oriented electric field reentrant resonator using two electronically tunable high Q hyperabrupt varactor diodes and feedback loops. In both moving phantoms and living mice, these automatic coupling control and automatic tuning control provisions resulted in an 8- to 10-fold increase in signal-to-noise ratio.     

Analysis of the tuning and operation of reflection resonator EPR spectrometers

This paper investigates basic characteristics of the electron paramagnetic resonance (EPR) signal obtained from spectrometers employing reflection resonators. General equations are presented which reveal the phase and amplitude dependence on instrumental parameters of both components of the continuous wave (CW) EPR signal (absorption and dispersion). New phase vector diagrams derived from these general equations are presented for the analysis of the EPR response. The dependence of the phase and absolute value of the CW EPR signal on the local oscillator (LO) phase and on resonator offset and coupling is presented and analyzed. The EPR spectrometer tuning procedures for both balanced and unbalanced heterodyne receivers are analyzed in detail using the new phase diagrams. Extraneous signals at the RF input of the microwave receiver (resulting from circulator leakage and reflections in the resonator transmission line) have been taken into account and analyzed. It is shown that a final tuning condition that corresponds to an extremum of the receiver output as a function of the resonator frequency is necessary and sufficient for the acquisition of pure absorption signal. This condition is universal: it applies to all spectrometer configurations in all frequency ranges. High Frequency EPR spectrometer (130 GHz) data are used to generate experimental phase diagrams that illustrate the theoretical concepts presented in the paper. Conditions are presented under which the absorption signal can be measured with complete suppression of the dispersion, independent of the mutual frequency offset between the microwave source and the EPR sample resonator. Equations describing the approximate relationship between changes of the resonator properties (Q-factor and frequency) and paramagnetic susceptibility are derived and analyzed.     

Electronically tunable surface-coil-type resonator for L-band EPR spectroscopy

The automatic frequency control (AFC) circuit in conventional electron paramagnetic resonance (EPR) spectrometers automatically tunes the microwave source to the resonance frequency of the resonator. The circuit works satisfactorily for samples stable enough that the geometric relations in the resonance structure do not change in a significant way. When EPR signals are measured during in vivo experiments with small rodents, however, the distance between the signal source and the surface-coil detector can change rapidly. When a conventional AFC circuit keeps the oscillator tuned to the resonator under those conditions, the resultant frequency change may exceed +/-5 MHz and markedly shift the position of the EPR signal. Such a shift results in unacceptable effects on the spectra, especially when the experimenter is dealing with narrow EPR lines. The animal movement also causes a mismatching of the resonator and the 50-ohm transmission line. Direct results of this mismatching are increased noise; shifts in the position of the baseline; and a high probability of overdriving the signal preamplifier with consequent loss of the EPR signal. We therefore designed, built, and tested a new surface-coil resonator using varactor diodes for tuning the resonance frequency to the fixed frequency oscillator and for capacitive matching of the resonator to the 50-ohm transmission line. The performance of the automatic matching system was tested in vivo by measuring EPR spectra of lithium phthalocyanine implanted in rats. Stability and sensitivity of the spectrometer were evaluated by measuring EPR spectra with and without the use of the automatic matching system. The overall experimental performance of the spectrometer was found to significantly improve during in vivo experiments using the automatic matching system. Excellent matching between the 50-ohm transmission line and the resonator was maintained under all experimental circumstances that were tested. This should allow us now to carry out experiments that previously were not possible.     

X-band rapid-scan EPR of nitroxyl radicals

X-band rapid-scan EPR spectra were obtained for dilute aqueous solutions of nitroxyl radicals (15)N-mHCTPO (4-hydro-3-carbamoyl-2,2,5,5-tetra-perdeuteromethyl-pyrrolin-1-(15)N-oxyl-d(12)) and (15)N-PDT (4-oxo-2,2,6,6-tetra-perdeuteromethyl-piperidinyl-(15)N-oxyl-d(16)). Simulations of spectra for (15)N-mHCTPO and (15)N-PDT agreed well with the experimental spectra. As the scan rate is increased in the rapid scan regime, the region in which signal amplitude increases linearly with B(1) extends to higher power and the maximum signal amplitude increases. In the rapid scan regime, the signal-to-noise for rapid-scan spectra was about a factor of 2 higher than for unbroadened CW EPR, even when the rapid scan spectra were obtained in a mode that had only 4% duty cycle for data acquisition. Further improvement in signal-to-noise per unit time is expected for higher duty cycles. Rapid scan spectra have higher bandwidth than CW spectra and therefore require higher detection bandwidths at faster scan rates. However, when the scan rate is increased by increasing the scan frequency, the increase in noise from the detection bandwidth is compensated by the decrease in noise due to increased number of averages per unit time. Because of the higher signal bandwidth, lower resonator Q is needed for rapid scan than for CW, so the rapid scan method is advantageous for lossy samples that inherently lower resonator Q.     

Single loop multi-gap resonator for whole body EPR imaging of mice at 1.2 GHz

For whole body EPR imaging of small animals, typically low frequencies of 250-750 MHz have been used due to the microwave losses at higher frequencies and the challenges in designing suitable resonators to accommodate these large lossy samples. However, low microwave frequency limits the obtainable sensitivity. L-band frequencies can provide higher sensitivity, and have been commonly used for localized in vivo EPR spectroscopy. Therefore, it would be highly desirable to develop an L-band microwave resonator suitable for in vivo whole body EPR imaging of small animals such as living mice. A 1.2 GHz 16-gap resonator with inner diameter of 42 mm and 48 mm length was designed and constructed for whole body EPR imaging of small animals. The resonator has good field homogeneity and stability to animal-induced motional noise. Resonator stability was achieved with electrical and mechanical design utilizing a fixed position double coupling loop of novel geometry, thus minimizing the number of moving parts. Using this resonator, high quality EPR images of lossy phantoms and living mice were obtained. This design provides good sensitivity, ease of sample access, excellent stability and uniform B(1) field homogeneity for in vivo whole body EPR imaging of mice at 1.2 GHz.     

Impact of the Characteristic Impedance of Coaxial Lines on the Sensitivity of a 750-MHz Electronically Tunable EPR Resonator

A 750-MHz electronically tunable resonator was investigated in terms of the sensitivity of electron paramagnetic resonance (EPR) signal detection. The conversion efficiency of the radio-frequency magnetic field was calculated for resonators with 50- and 100-Ω coaxial coupling lines using three-dimensional (3D) microwave field and microwave circuit simulators. Based on the simulation results, two tunable resonators were physically constructed and compared in terms of EPR signal sensitivity using a nitroxyl radical solution. While the resonator with 100-Ω coaxial lines provided 14% greater signal intensity, its signal-to-noise ratio was lower than that of the resonator with 50-Ω lines. To demonstrate the capability of the constructed tunable resonator for EPR imaging experiments, a solution of nitroxyl radical and the leg of a tumor-bearing mouse were visualized.     

A bridged loop-gap S-band surface resonator for topical EPR spectroscopy.

The design and structure of a bridged loop-gap surface resonator developed for topical EPR spectroscopy and imaging of the distribution and metabolism of spin labels in in vivo skin is reported. The resonator is a one-loop, one-gap bridged structure. A pivoting single loop-coupling coil was used to couple the microwave power to the loop-gap resonant structure. A symmetric coupling circuit was used to achieve better shielding and minimize radiation. The frequency of the resonator can be easily adjusted by trimming the area of the capacitive foil bridge, which overlaps the gap in the cylindrical loop. The working frequency set was 2.2 GHz and the unloaded Q was 720. The B1 field of this resonator was measured and spatially mapped by three-dimensional EPR imaging. The resonator is well suited to topical measurements of large biological subjects and is readily applicable for in vivo measurements of free radicals in human skin.     

A tunable general purpose Q-band resonator for CW and pulse EPR/ENDOR experiments with large sample access and optical excitation

We describe a frequency tunable Q-band cavity (34 GHz) designed for CW and pulse Electron Paramagnetic Resonance (EPR) as well as Electron Nuclear Double Resonance (ENDOR) and Electron Electron Double Resonance (ELDOR) experiments. The TE(011) cylindrical resonator is machined either from brass or from graphite (which is subsequently gold plated), to improve the penetration of the 100 kHz field modulation signal. The (self-supporting) ENDOR coil consists of four 0.8mm silver posts at 2.67 mm distance from the cavity center axis, penetrating through the plunger heads. It is very robust and immune to mechanical vibrations. The coil is electrically shielded to enable CW ENDOR experiments with high RF power (500 W). The top plunger of the cavity is movable and allows a frequency tuning of ±2 GHz. In our setup the standard operation frequency is 34.0 GHz. The microwaves are coupled into the resonator through an iris in the cylinder wall and matching is accomplished by a sliding short in the coupling waveguide. Optical excitation of the sample is enabled through slits in the cavity wall (transmission ～60%). The resonator accepts 3mm o.d. sample tubes. This leads to a favorable sensitivity especially for pulse EPR experiments of low concentration biological samples. The probehead dimensions are compatible with that of Bruker flexline Q-band resonators and it fits perfectly into an Oxford CF935 Helium flow cryostat (4-300 K). It is demonstrated that, due to the relatively large active sample volume (20-30 μl), the described resonator has superior concentration sensitivity as compared to commercial pulse Q-band resonators. The quality factor (Q(L)) of the resonator can be varied between 2600 (critical coupling) and 1300 (over-coupling). The shortest achieved π/2-pulse durations are 20 ns using a 3 W microwave amplifier. ENDOR (RF) π-pulses of 20 μs ((1)H @ 51 MHz) were obtained for a 300 W amplifier and 7 μs using a 2500 W amplifier. Selected applications of the resonator are presented.     

Generating EPR-entangled mechanical state via feeding finite-bandwidth squeezed light

Einstein–Podolski–Rosen(EPR) entanglement state is achievable by combining two single-mode position and momentum squeezed states at a 50:50 beam-splitter(BS). We investigate the generation of the EPR entangled state of two vibrating membranes in a ring resonator, where clockwise(CW) and counter-clockwise(CCW) travelling-wave modes are driven by lasers and finite-bandwidth squeezed lights. Since the optomechanical coupling depends on the location of the membranes, CW and CCW can couple to the symmetric and antisymmetric combination of mechanical modes for a suitable arrangement, which corresponds to a 50:50 BS mixing. Moreover, by employing the red-detuned driving laser and tuning the central frequency of squeezing field blue detuned from the driving laser with a mechanical frequency, the squeezing property of squeezed light can be perfectly transferred to the mechanical motion in the weak coupling regime. Thus, the BS mixing modes can be position and momentum squeezed by feeding the appropriate squeezed lights respectively, and the EPR entangled mechanical state is obtained. Moreover, cavity-induced mechanical cooling can further suppress the influence of thermal noise on the entangled state.     

SOI微环谐振腔测试信号纹波噪声的分析

基于绝缘体上硅材料（SOI）的微环谐振腔作为各种集成微光学器件的核心部件,其传输特性的好坏直接决定微光学器件的性能优劣。结合微环谐振腔理论和多光束干涉原理对微环谐振腔测试信号中纹波的产生原因和影响因素进行了分析,通过耦合实验测试得到了纹波信号的特征参数并进行了分析,实验结果与仿真分析吻合。进一步分析了耦合光栅刻蚀深度对纹波幅值的影响及原因,为微环谐振腔结构参数的进一步优化和微环谐振腔传感系统误差分析提供了依据。     

In vivo temporal EPR study using a region-selected intensity determination method to estimate cerebral reducing ability in rats treated with olanzapine

Region-selected intensity determination (RSID) is a method for obtaining the temporal changes in electron paramagnetic resonance (EPR) signal intensity from a target region, without the use of complicated procedures employed in the conventional imaging methods. An in vivo 700-MHz radio frequency EPR spectrometer equipped with a bridged loop-gap resonator was used with the RSID method to estimate intracerebral reducing ability in the rat following acute administration of olanzapine (OZP) or haloperidol (HPD). To this end, temporal changes in EPR signal intensity of target regions (the striatum and the prefrontal cortex) of rats which had received a blood-brain-barrier-permeable nitroxide radical (3-hydroxymethyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl) via an intravenous route were observed. The half-lives of EPR signal intensity in both regions of OZP- or HPD-treated rats were significantly longer than in control animals. This indicated that reducing abilities of the striatum and cerebral cortex decreased in the rats to which either OZP or HPD had been acutely administered.     

Digital EPR with an arbitrary waveform generator and direct detection at the carrier frequency

A digital EPR spectrometer was constructed by replacing the traditional bridge with an arbitrary waveform generator (AWG) to produce excitation patterns and a high-speed digitizer for direct detection of the spin system response at the carrier frequency. Digital down-conversion produced baseband signals in quadrature with very precise orthogonality. Real-time resonator tuning was performed by monitoring the Fourier transforms of signals reflected from the resonator during frequency sweeps generated by the AWG. The capabilities of the system were demonstrated by rapid magnetic field scans at 256 MHz carrier frequency, and FID and spin echo experiments at 1 and 10 GHz carrier frequencies. For the rapid scan experiments the leakage through a cross-loop resonator was compensated by adjusting the amplitude and phase of a sinusoid at the carrier frequency that was generated with another AWG channel.     

Systematic approach to cutoff frequency selection in continuous-wave electron paramagnetic resonance imaging

This article describes a systematic method for determining the cutoff frequency of the low-pass window function that is used for deconvolution in two-dimensional continuous-wave electron paramagnetic resonance (EPR) imaging. An evaluation function for the criterion used to select the cutoff frequency is proposed, and is the product of the effective width of the point spread function for a localized point signal and the noise amplitude of a resultant EPR image. The present method was applied to EPR imaging for a phantom, and the result of cutoff frequency selection was compared with that based on a previously reported method for the same projection data set. The evaluation function has a global minimum point that gives the appropriate cutoff frequency. Images with reasonably good resolution and noise suppression can be obtained from projections with an automatically selected cutoff frequency based on the present method.     

Parallel coil resonators for time-domain radiofrequency electron paramagnetic resonance imaging of biological objects

Resonators suitable for time-domain electron paramagnetic resonance spectroscopy and imaging at a radiofrequency capable of accommodating experimental animals such as mice are described. Design considerations included B(1) field homogeneity, optimal Q, spectral bandwidth, resonator ring-down, and sensitivity. Typically, a resonator with 25-mm diameter and 25-mm length was constructed by coupling 11 single loops in parallel with a separation of 2.5 mm. To minimize the resonator ring-down time and provide the necessary spectral bandwidth for in vivo imaging experiments, the Q was reduced predominantly by overcoupling. Capacitative coupling was utilized to minimize microphonic effects. The B(1) field in the resonator was mapped both radially and axially and found to be uniform and adequate for imaging studies. Imaging studies with phantom objects containing a narrow-line spin probe as well as in vivo objects administered with the spin probe show the suitability of these resonators for valid reproduction of the spin probe distribution in three dimensions. The fabrication of such resonators is simple and can be scaled up with relative ease to accommodate larger objects as well.     

基于滑模极值搜索的SSC-LINAC RFQ调谐系统设计与测试

将滑模极值搜索算法引入基于腔体反射信号的SSC-LINAC射频四极场加速器(Radio Frequency Quadrupole,RFQ)腔体频率稳定系统设计中,结合Lyapunov稳定性理论和SSC-LINAC RFQ高频系统的特点,对滑模极值搜索算法的控制增益和参考信号进行改进;通过求解微分方程,得到腔体反射信号关...