Fast 3D spatial EPR imaging using spiral magnetic field gradient

Electron paramagnetic resonance imaging (EPRI) provides direct detection and mapping of free radicals. The continuous wave (CW) EPRI technique, in particular, has been widely used in a variety of applications in the fields of biology and medicine due to its high sensitivity and applicability to a wide range of free radicals and paramagnetic species. However, the technique requires long image acquisition periods, and this limits its use for many in vivo applications where relatively rapid changes occur in the magnitude and distribution of spins. Therefore, there has been a great need to develop fast EPRI techniques. We report the development of a fast 3D CW EPRI technique using spiral magnetic field gradient. By spiraling the magnetic field gradient and stepping the main magnetic field, this approach acquires a 3D image in one sweep of the main magnetic field, enabling significant reduction of the imaging time. A direct one-stage 3D image reconstruction algorithm, modified for reconstruction of the EPR images from the projections acquired with the spiral magnetic field gradient, was used. We demonstrated using a home-built L-band EPR system that the spiral magnetic field gradient technique enabled a 4-7-fold accelerated acquisition of projections. This technique has great potential for in vivo studies of free radicals and their metabolism.     

Object dependent sweep width reduction with spectral-spatial EPR imaging

For spectral-spatial EPR imaging, prior knowledge about the spatial support of an imaged object can be exploited in two ways. We can shrink the spatial field of view (FOV) to closely wrap the object in a sphere or reduce the sweep width in a projection dependent fashion. Use of a smaller spatial FOV with the same number of samples enhances spatial resolution by reducing voxel volume at the expense of signal-to-noise and a consequent degraded line-width resolution. We have developed another approach to define sweep width that prunes away the portions of the projection sweep with no signal. This reduces data acquisition time for the continuous wave (CW) EPR image proportional to the sweep width reduction. This method also avoids voxel volume reduction. Using the reduced-sweep method, we decreased the data acquisition time by 20% maintaining spatial and linewidth resolution.     

Electron paramagnetic resonance imaging using magnetic-field-gradient spinning

A novel X-band CW EPR imaging has been developed using magnetic-field-gradient (MFG) spinning to obtain spatial distributions of electron paramagnetic species. Spinning MFG EPR imaging for 65 projection spectra required just 55 s while conventional imaging took 11 min 40 s, that is, the acquisition time for the new system is one order of magnitude shorter than that for conventional EPR imaging. Spinning MFG EPR imaging allows one to measure reconstructed images in an interactive manner where resolution and condition can be changed quickly.     

Evaluation and comparison of pulsed and continuous wave radiofrequency electron paramagnetic resonance techniques for in vivo detection and imaging of free radicals

The performance of two electron paramagnetic resonance (EPR) spectrometers/imagers, one configured in pulsed mode and the other in continuous wave (CW) mode, at an operating frequency of 300 MHz is compared. Using the same resonator (except for altered Q-factors), identical samples and filling factors in the two techniques have been evaluated for their potentials and limitations for in vivo spectroscopic and imaging applications. The assessment is based on metrics such as sensitivity, spatial and temporal resolution, field of view, image artifacts, viable spin probes, and subjects of study. The spectrometer dead time limits the pulsed technique to samples with long phase memories (>275 ns). Nevertheless, for viable narrow-line spin probes, the pulsed technique offers better sensitivity and temporal resolution. The CW technique, on the other hand, does not restrict the choice at spin probes. In addition, the phase-sensitive narrow-band detection of the CW technique gives artifact-free images even for large objects. Selected examples illustrating the performance of the CW and pulsed techniques are presented to put the capabilities of the two techniques in perspective.     

Biochemical EPR imaging of skin

EPR Imaging (EPRI) of spin labels is a powerful method for investigating skin and can give information about biochemical processes which are involved in numerous skin diseases. Furthermore it enables the non invasive investigation of the liberation, penetration and distribution of spin labelled drugs. The basis of these measurements is spectral spatial EPR imaging employing modulated field gradients and simultaneous field scans (MOSS). A skin region (?=6 mm) was treated with a 10 μl spin label solution (1 mM). EPR spectra of 128 points were recorded in 128 spatial planes resulting in a 128×128 image matrix. A spatial resolution of better than 10 μm can be obtained for a spectral line width of 0.1 mT and a gradient of 4 Tm^?1.In vivo imaging on mammalian skin can be performed by employing surface coils at S-band frequencies, 3 GHz.     

Simple data acquisition method for multi-dimensional EPR spectral-spatial imaging using a combination of constant-time and projection-reconstruction modalities

A combination of the constant-time spectral-spatial imaging (CTSSI) modality and projection-reconstruction modality was tested to simplify data acquisition for multi-dimensional CW EPR spectral-spatial imaging. In this method, 3D spectral-spatial image data were obtained by simple repetition of conventional 2D CW imaging process, except that the field gradient amplitude was incremented in constant steps in each repetition. The data collection scheme was no different from the conventional CW imaging system for spectral-spatial data acquisition. No special equipment and/or rewriting of existing software were required. The data acquisition process for multi-dimensional spectral-spatial imaging is consequently simplified. There is also no “missing-angle” issue because the CTSSI modality was employed to reconstruct 2D spectral-spatial images. Extra reconstruction processes to obtain higher spatial dimensions were performed using a conventional projection-reconstruction modality. This data acquisition technique can be applied to any conventional CW EPR (spatial) imaging system for multi-dimensional spectral-spatial imaging.     

Detection of undistorted continuous wave (CW) electron paramagnetic resonance (EPR) spectra with non-adiabatic rapid sweep (NARS) of the magnetic field

A continuous wave (CW) electron paramagnetic resonance (EPR) spectrum is typically displayed as the first harmonic response to the application of 100 kHz magnetic field modulation, which is used to enhance sensitivity by reducing the level of 1/f noise. However, magnetic field modulation of any amplitude causes spectral broadening and sacrifices EPR spectral intensity by at least a factor of two. In the work presented here, a CW rapid-scan spectroscopic technique that avoids these compromises and also provides a means of avoiding 1/f noise is developed. This technique, termed non-adiabatic rapid sweep (NARS) EPR, consists of repetitively sweeping the polarizing magnetic field in a linear manner over a spectral fragment with a small coil at a repetition rate that is sufficiently high that receiver noise, microwave phase noise, and environmental microphonics, each of which has 1/f characteristics, are overcome. Nevertheless, the rate of sweep is sufficiently slow that adiabatic responses are avoided and the spin system is always close to thermal equilibrium. The repetitively acquired spectra from the spectral fragment are averaged. Under these conditions, undistorted pure absorption spectra are obtained without broadening or loss of signal intensity. A digital filter such as a moving average is applied to remove high frequency noise, which is approximately equivalent in bandwidth to use of an integrating time constant in conventional field modulation with lock-in detection. Nitroxide spectra at L- and X-band are presented.     

Kinetic measurements using EPR imaging with a modulated field gradient

EPR imaging with modulated field gradient was applied for the investigation of fast diffusion processes. Three different imaging methods are possible: spectral-temporal, spatio-temporal, and spectral-spatial imaging. The time resolution is on the order of seconds and the spatial resolution is in the micrometer region. The efficiency of this imaging technique is demonstrated for the penetration of the spin probe Tempol in the skin of hairless mice biopsies. The skin is normally protected against the penetration of water soluble substances by the horny layer, a resistive thin lipophilic layer. Overcoming this horny layer for water soluble ingredients is one of the main practical problems for the topical application of pharmaceutics which could be investigated by EPR imaging. Different images represent the penetration behavior of the water soluble Tempol in the skin after treatment with the penetration enhancer DMSO (Dimethylsulfoxide) and after removing the horny layer.     

Variable radio frequency proton-electron double-resonance imaging: application to pH mapping of aqueous samples

Proton-electron double-resonance imaging (PEDRI) offers rapid image data collection and high resolution for spatial distribution of paramagnetic probes. Recently we developed the concept of variable field (VF) PEDRI which enables extracting a functional map from a limited number of images acquired at pre-selected EPR excitation fields using specific paramagnetic probes (Khramtsov et al., J. Magn. Reson. 202 (2010) 267-273). In this work, we propose and evaluate a new modality of PEDRI-based functional imaging with enhanced temporal resolution which we term variable radio frequency (VRF) PEDRI. The approach allows for functional mapping (e.g., pH mapping) using specifically designed paramagnetic probes with high quality spatial resolution and short acquisition times. This approach uses a stationary magnetic field but different EPR RFs. The ratio of Overhauser enhancements measured at each pixel at two different excitation frequencies corresponding to the resonances of protonated and deprotonated forms of a pH-sensitive nitroxide is converted to a pH map using a corresponding calibration curve. Elimination of field cycling decreased the acquisition time by exclusion periods of ramping and stabilization of the magnetic field. Improved magnetic field homogeneity and stability allowed for the fast MRI acquisition modalities such as fast spin echo. In total, about 30-fold decrease in EPR irradiation time was achieved for VRF PEDRI (2.4s) compared with VF PEDRI (70s). This is particularly important for in vivo applications enabling one to overcome the limiting stability of paramagnetic probes and sample overheating by reducing RF power deposition.     

A parametric approach to spectral-spatial EPR imaging

Continuous wave electron paramagnetic resonance imaging for in vivo mapping of spin distribution and spectral shape requires rapid data acquisition. A spectral-spatial imaging technique is presented that provides an order of magnitude reduction in acquisition time, compared to iterative tomographic reprojection. The proposed approach assumes that spectral shapes in the sample are well-approximated by members from a parametric family of functions. A model is developed for the spectra measured with magnetic field modulation. Parameters defining the spin distribution and spectral shapes are then determined directly from the measurements using maximum a posteriori probability estimation. The approach does not suffer approximation error from limited sweep width of the main magnetic field and explicitly incorporates the variability in signal-to-noise ratio versus strength of magnetic field gradient. The processing technique is experimentally demonstrated on a one-dimensional phantom containing a nitroxide spin label with constant g-factor. Using an L-band EPR spectrometer, spectral shapes and spin distribution are accurately recovered from two projections and a spectral window which is comparable to the maximum linewidth of the sample.     

Time-of-flight flow imaging using NMR remote detection

A time-of-flight imaging technique is introduced to visualize fluid flow and dispersion through porous media using NMR. As the fluid flows through a sample, the nuclear spin magnetization is modulated by rf pulses and magnetic field gradients to encode the spatial coordinates of the fluid. When the fluid leaves the sample, its magnetization is recorded by a second rf coil. This scheme not only facilitates a time-dependent imaging of fluid flow, it also allows a separate optimization of encoding and detection subsystems to enhance overall sensitivity. The technique is demonstrated by imaging gas flow through a porous rock.     

Solving the problem of concomitant gradients in ultra-low-field MRI

In ultra-low-field magnetic resonance imaging (ULF MRI), spin precession is detected typically in magnetic fields of the order of 10-100 μT. As in conventional high-field MRI, the spatial origin of the signals can be encoded by superposing gradient fields on a homogeneous main field. However, because the main field is weak, gradient field amplitudes become comparable to it. In this case, the concomitant gradients forced by Maxwell's equations cause the assumption of linearly varying field gradients to fail. Thus, image reconstruction with Fourier transformation would produce severe image artifacts. We propose a direct linear inversion (DLI) method to reconstruct images without limiting assumptions about the gradient fields. We compare the quality of the images obtained using the proposed reconstruction method and the Fourier reconstruction. With simulations, we show how the reconstruction errors of the methods depend on the strengths of the concomitant gradients. The proposed approach produces nearly distortion-free images even when the main field reaches zero.     

Sensitivity quantification of remote detection NMR and MRI

A sensitivity analysis is presented of the remote detection NMR technique, which facilitates the spatial separation of encoding and detection of spin magnetization. Three different cases are considered: remote detection of a transient signal that must be encoded point-by-point like a free induction decay, remote detection of an experiment where the transient dimension is reduced to one data point like phase encoding in an imaging experiment, and time-of-flight (TOF) flow visualization. For all cases, the sensitivity enhancement is proportional to the relative sensitivity between the remote detector and the circuit that is used for encoding. It is shown for the case of an encoded transient signal that the sensitivity does not scale unfavorably with the number of encoded points compared to direct detection. Remote enhancement scales as the square root of the ratio of corresponding relaxation times in the two detection environments. Thus, remote detection especially increases the sensitivity of imaging experiments of porous materials with large susceptibility gradients, which cause a rapid dephasing of transverse spin magnetization. Finally, TOF remote detection, in which the detection volume is smaller than the encoded fluid volume, allows partial images corresponding to different time intervals between encoding and detection to be recorded. These partial images, which contain information about the fluid displacement, can be recorded, in an ideal case, with the same sensitivity as the full image detected in a single step with a larger coil.     

Spin relaxation measurements using first-harmonic out-of-phase absorption EPR signals: rotational motion effects

A recent survey of nonlinear continuous-wave (CW) EPR methods revealed that the first-harmonic absorption EPR signal, detected 90 degrees out of phase with respect to the Zeeman modulation (V(1)(')-EPR), is the most appropriate for determining spin-lattice relaxation enhancements of spin labels (V. A. Livshits, T. Páli, and D. Marsh, 1998, J. Magn. Reson. 134, 113-123). The sensitivity of such V(1)(')-EPR spectra to molecular rotational motion is investigated here by spectral simulations for nitroxyl spin labels, over the entire range of rotational correlation times. Determination of the effective spin-lattice relaxation times is less dependent on rotational mobility than for other nonlinear CW EPR methods, especially at a Zeeman modulation frequency of 25 kHz which is particularly appropriate for spin labels. This relative insensitivity to molecular motion further enhances the usefulness of the V(1)(')-EPR method. Calibrations of the out-of-phase to in-phase spectral intensity (and amplitude) ratios are given as a function of spin-lattice relaxation time, for the full range of spin-label rotational correlation times. Experimental measurements on spin labels in the slow, intermediate, and fast motional regimes of molecular rotation are used to test and validate the method.     

Comparison of Continuous Wave, Spin Echo, and Rapid Scan EPR of Irradiated Fused Quartz

The E' defect in irradiated fused quartz has spin lattice relaxation times (T(1)) about 100 to 300 μs and spin-spin relaxation times (T(2)) up to about 200 μs, depending on the concentration of defects and other species in the sample. These long relaxation times make it difficult to record an unsaturated continuous wave (CW) electron paramagnetic resonance (EPR) signal that is free of passage effects. Signals measured at X-band (~9.5 GHz) by three EPR methods: conventional slow-scan field modulated EPR, rapid scan EPR, and pulsed EPR, were compared. To acquire spectra with comparable signal-to-noise, both pulsed and rapid scan EPR require less time than conventional CW EPR. Rapid scan spectroscopy does not require the high power amplifiers that are needed for pulsed EPR. The pulsed spectra, and rapid scan spectra obtained by deconvolution of the experimental data, are free of passage effects.     

Resonant-Optical Detection of Nuclear Magnetization in the Si/CaF<Subscript>2</Subscript> Nanostructure

We have considered theoretically the characteristic features of optical detection of nuclear magnetization in the Si/CaF₂ structure under the conditions of EPR and NMR by measuring luminescence polarization. We show that application of EPR makes it possible to detect weak nuclear fields, but the time of spin relaxation of electrons imposes its constraint. The application of NMR is limited by nuclear fields of no less than 15–20 Gs. The possibility of using optical NMR for direct measurement of the nuclear field from changes in the luminescence polarization spectrum is shown. __________ Translated from Zhurnal Prikladnoi Spektroskopii, Vol. 72, No. 5, pp. 644–649, September–October, 2005.     

Mapping the B1 field distribution with nonideal gradients in a high-resolution NMR spectrometer

To understand the behavior of many NMR experiments, it is important to determine the spatial distribution of the B1 field. In this paper, we show how this distribution can be mapped independently of spin density, coil responsiveness, and nonlinearities of the B0 field gradients. As a by-product we obtain a map of the (possibly nonlinear) spatial variation of the B0 field gradients used in the imaging procedure.     

EPR oximetry in three spatial dimensions using sparse spin distribution

A method is presented to use continuous wave electron paramagnetic resonance imaging for rapid measurement of oxygen partial pressure in three spatial dimensions. A particulate paramagnetic probe is employed to create a sparse distribution of spins in a volume of interest. Information encoding location and spectral linewidth is collected by varying the spatial orientation and strength of an applied magnetic gradient field. Data processing exploits the spatial sparseness of spins to detect voxels with nonzero spin and to estimate the spectral linewidth for those voxels. The parsimonious representation of spin locations and linewidths permits an order of magnitude reduction in data acquisition time, compared to four-dimensional tomographic reconstruction using traditional spectral-spatial imaging. The proposed oximetry method is experimentally demonstrated for a lithium octa-n-butoxy naphthalocyanine (LiNc–BuO) probe using an L-band EPR spectrometer.     

Echo-planar rotating-frame imaging

A new rotating-frame imaging method that produces a complete cross section of an object in a single experiment is reported. The echo planar rotating frame imaging (EPROFI) technique uses two perpendicular RF gradients for two-dimensional spatial encoding and fully exploits the formation of rotary echoes for fast sampling of spatial frequencies. The acquisition scheme yields the Fourier transform of the spin distribution on Cartesian coordinates for straightforward image reconstruction. Implementation of the technique on a low-field portable NMR probe is described and results are presented for test objects with different geometries.