Overhauser-enhanced magnetic resonance elastography.

Magnetic resonance elastography (MRE) is a powerful technique to assess the mechanical properties of living tissue. However, it suffers from reduced sensitivity in regions with short T2 and T2 * such as in tissue with high concentrations of paramagnetic iron, or in regions surrounding implanted devices. In this work, we exploit the longer T2 * attainable at ultra-low magnetic fields in combination with Overhauser dynamic nuclear polarization (DNP) to enable rapid MRE at 0.0065 T. A 3D balanced steady-state free precession based MRE sequence with undersampling and fractional encoding was implemented on a 0.0065 T MRI scanner. A custom-built RF coil for DNP and a programmable vibration system for elastography were developed. Displacement fields and stiffness maps were reconstructed from data recorded in a polyvinyl alcohol gel phantom loaded with stable nitroxide radicals. A DNP enhancement of 25 was achieved during the MRE sequence, allowing the acquisition of 3D Overhauser-enhanced MRE (OMRE) images with (1.5 × 2.7 × 9) mm(3) resolution over eight temporal steps and 11 slices in 6 minutes. In conclusion, OMRE at ultra-low magnetic field can be used to detect mechanical waves over short acquisition times. This new modality shows promise to broaden the scope of conventional MRE applications, and may extend the utility of low-cost, portable MRI systems to detect elasticity changes in patients with implanted devices or iron overload.

Two-dimensional imaging in a lightweight portable MRI scanner without gradient coils.

PURPOSE: As the premiere modality for brain imaging, MRI could find wider applicability if lightweight, portable systems were available for siting in unconventional locations such as intensive care units, physician offices, surgical suites, ambulances, emergency rooms, sports facilities, or rural healthcare sites. METHODS: We construct and validate a truly portable (<100 kg) and silent proof-of-concept MRI scanner which replaces conventional gradient encoding with a rotating lightweight cryogen-free, low-field magnet. When rotated about the object, the inhomogeneous field pattern is used as a rotating spatial encoding magnetic field (rSEM) to create generalized projections which encode the iteratively reconstructed two-dimensional (2D) image. Multiple receive channels are used to disambiguate the nonbijective encoding field. RESULTS: The system is validated with experimental images of 2D test phantoms. Similar to other nonlinear field encoding schemes, the spatial resolution is position dependent with blurring in the center, but is shown to be likely sufficient for many medical applications. CONCLUSION: The presented MRI scanner demonstrates the potential for portability by simultaneously relaxing the magnet homogeneity criteria and eliminating the gradient coil. This new architecture and encoding scheme shows convincing proof of concept images that are expected to be further improved with refinement of the calibration and methodology.

High speed 3D overhauser-enhanced MRI using combined b-SSFP and compressed sensing.

PURPOSE: Overhauser-enhanced MRI is a promising technique for imaging the distribution and dynamics of free radicals. A key challenge for Overhauser-enhanced MRI is attaining high spatial and temporal resolution while simultaneously limiting resonator and sample heating due to the long, high power radio-frequency pulses needed to saturate the electron resonance. METHODS: The approach presented here embeds EPR pulses within a balanced steady state free precession sequence. Unlike other Overhauser-enhanced MRI methods, no separate Overhauser prepolarization step is required. This steady-state approach also eliminates the problem of time-varying Overhauser-enhanced signal and provides constant polarization in the sample during the acquisition. A further increase in temporal resolution was achieved by incorporating undersampled k-space strategies and compressed sensing reconstruction. RESULTS: We demonstrate 1 × 2 × 3.5 mm(3) resolution at 6.5 mT across a 54 × 54 × 110 mm(3) sample in 33 s while sampling 30% of k-space. CONCLUSION: The work presented here overcomes the main limitations of Overhauser enhanced MRI as previously described in the literature, drastically improving speed and resolution, and enabling new opportunities for the measurement of free radicals in living organisms, and for the study of dynamic processes such as metabolism and flow.

Temperature dependence of high field 13C dynamic nuclear polarization processes with trityl radicals below 35 Kelvin.

In order to facilitate versatile applications with high field dynamic nuclear polarization (DNP), it is important to be able to optimize the DNP performance, i.e. reach high nuclear hyperpolarization within a short signal build up time. Given that the solid-state DNP process is strongly temperature-dependent, it is important to benchmark the temperature dependence of various DNP and electron paramagnetic resonance (EPR) parameters that can then be used to test and develop theories and models for high field DNP mechanisms. However, DNP and EPR experiments at high fields and cryogenic temperatures below 20 Kelvin usually require home built instrumentation, and therefore even basic experimental observations are lacking in the literature. DNP and EPR experiments at 7 T (197 GHz) and 8.5 T (240 GHz), respectively, were conducted at temperatures between 35 K and 3.7 K where the electron thermal polarization changes from 13.4% to 85.6%, respectively. The samples are frozen solutions of 15 mM OX063Me trityl radicals in various mixtures of [1-(13)C]pyruvic acid, glycerol, and Gd(3+)-chelates. For all sample mixtures, the trityl EPR lines are found to be inhomogeneously broadened and the dominant DNP mechanism is shown to be the cross effect (CE). A 20%, 11%, and 6.77% (13)C polarization is achieved at 3.7 K with a [1-(13)C]pyruvic-glycerol-H2O sample, the addition of 2 mM of Gd(3+)-chelates, and pure [1-(13)C]pyruvic acid, respectively. When T1n is sufficiently long, our results seem to suggest T1e is a key variable in the DNP process, where longer T1e values correlate with larger DNP enhancements (εDNP). The experimental data reported here on the temperature dependence of T1n, T1e, Tm (electron phase memory time), the EPR linewidth, TDNP and ε(DNP) at high fields will be helpful for testing the mechanism and theory of DNP processes.

Inductively coupled NMR probe for versatile dynamic nuclear polarization operation at 7 T: observation of 61 ± 2% ¹H polarization at 4 K.

We have performed dynamic nuclear polarization (DNP) experiments at liquid helium temperatures using a low-power (<70 mW) solid-state diode microwave source at 200 GHz-the electron paramagnetic resonance frequency of stable radicals at 7 T. We employed a home-built Alderman-Grant probe for the detection of ¹H NMR signal at 300 MHz, as such coils are well suited for higher frequency NMR detection. The Alderman-Grant coil is inductively coupled to the rest of the radiofrequency (rf) circuit, whose design allows probe components to be placed away from the sample area, and also enables easy switching of coils with different diameters and resonance frequencies. We have tested our DNP instrument on a frozen nitroxide model system consisting of 4-Amino TEMPO dissolved in a glycerol:water mixture. The largest nuclear spin polarization observed was 61 ± 2% with a sample containing 20 mM 4-Amino TEMPO dissolved in deuterated glycerol (d-glycerol):D2O:H2O (50:40:10), amounting to record polarization measured to date at an easily amenable temperature of 4 K.

Site-specific hydration dynamics in the nonpolar core of a molten globule by dynamic nuclear polarization of water.

Water-protein interactions play a direct role in protein folding. The chain collapse that accompanies protein folding involves extrusion of water from the nonpolar core. For many proteins, including apomyoglobin (apoMb), hydrophobic interactions drive an initial collapse to an intermediate state before folding to the final structure. However, the debate continues as to whether the core of the collapsed intermediate state is hydrated and, if so, what the dynamic nature of this water is. A key challenge is that protein hydration dynamics is significantly heterogeneous, yet suitable experimental techniques for measuring hydration dynamics with site-specificity are lacking. Here, we introduce Overhauser dynamic nuclear polarization at 0.35 T via site-specific nitroxide spin labels as a unique tool to probe internal and surface protein hydration dynamics with site-specific resolution in the molten globular, native, and unfolded protein states. The (1)H NMR signal enhancement of water carries information about the local dynamics of the solvent within ∼10 Šof a spin label. EPR is used synergistically to gain insights on local polarity and mobility of the spin-labeled protein. Several buried and solvent-exposed sites of apoMb are examined, each bearing a covalently bound nitroxide spin label. We find that the nonpoloar core of the apoMb molten globule is hydrated with water bearing significant translational dynamics, only 4-6-fold slower than that of bulk water. The hydration dynamics of the native state is heterogeneous, while the acid-unfolded state bears fast-diffusing hydration water. This study provides a high-resolution glimpse at the folding-dependent nature of protein hydration dynamics.

A 200 GHz dynamic nuclear polarization spectrometer.

We present our experimental setup for both dynamic nuclear polarization (DNP) and electron paramagnetic resonance (EPR) detection at 7 T using a quasi-optical bridge for propagation of the 200 GHz beam and our initial results obtained at 4 K. Our quasi-optical bridge allows the polarization of the microwave beam to be changed from linear to circular. Only the handedness of circular polarization in the direction of the Larmor precession is absorbed by the electron spins, so a gain in effective microwave power of two is expected for circular vs. linear polarization. Our results show an increase in DNP signal enhancement of 28% when using circularly vs. linearly polarized radiation. We measured a maximum signal enhancement of 65 times that of thermal polarization for a (13)C labeled urea sample corresponding to 3% nuclear spin polarization. Since the time constant for nuclear spin polarization buildup during microwave irradiation is 10 times faster than the (13)C nuclear spin T(1), the actual gain in detection sensitivity with DNP is much greater.

Overhauser dynamic nuclear polarization and molecular dynamics simulations using pyrroline and piperidine ring nitroxide radicals.

The efficiency of Overhauser dynamic nuclear polarization (DNP) depends on the local dynamics modulating the dipolar coupling between the two interacting spins. By attaching nitroxide based spin labels to molecules and by measuring the (1)H DNP response of solvent water, information about the local hydration dynamics near the spin label can be obtained. However, there are two commonly used types of nitroxide ring structures; a pyrroline based and a piperidine based molecule. It is important to know when comparing different experiments, whether changes in DNP enhancements are due to changes in local hydration dynamics or because of the different spin label structures. In this study we investigate the key parameters affecting DNP signal enhancements for 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrrolin-1-oxyl, a 5-membered ring nitroxide radical, and for 4-oxo-2,2,6,6-tetramethyl-1-piperidinyloxy, a 6-membered ring nitroxide radical. Using X-Band DNP, field cycling relaxometry, and molecular dynamics simulations, we conclude that the key parameters affecting the DNP amplitude of the (1)H signal of water to be equal when using either nitroxide. Thus, experiments measuring hydration dynamics using either type of spin labels may be compared.

Overhauser dynamic nuclear polarization to study local water dynamics.

Surface and internal water dynamics of molecules and soft matter are of great relevance to their structure and function, yet the experimental determination under ambient and steady-state conditions is challenging. One of the most powerful approaches to measure local water dynamics within 5 A distances is to utilize the modulation of the nuclear spin relaxation rate of water protons through their time-dependent dipolar coupling to paramagnetic probes, here nitroxide spin labels. We recently introduced a method to obtain local water dynamics through Overhauser dynamic nuclear polarization (DNP). This has a unique advantage over other related techniques available in that a highly amplified proton nuclear magnetic resonance signal carries the information, allowing the use of minute microliter sample volumes and 100 muM sample concentrations. The outcome of our approach is the quantitative determination of the key DNP parameter known as the coupling factor, which provides local translational diffusion dynamics of the solvent within 5 A of the spin label. In contrast to recent reports that the coupling factor for nitroxide radicals cannot be quantified due to the difficulty in determining the saturation factor for the spin label, we show the saturation factor can be accurately determined and for the first time present agreement between measurements and theory. We discuss the discrepancy between the related field cycling relaxometery technique and DNP in determining the coupling factor and present arguments in support of the DNP-determined value. DNP measurements of local hydration dynamics around nitroxides in bulk water and on the surface of proteins are presented.

Dynamic nuclear polarization enhanced nuclear magnetic resonance and electron spin resonance studies of hydration and local water dynamics in micelle and vesicle assemblies.

We present a unique analysis tool for the selective detection of local water inside soft molecular assemblies (hydrophobic cores, vesicular bilayers, and micellar structures) suspended in bulk water. Through the use of dynamic nuclear polarization (DNP), the (1)H NMR signal of water is amplified, as it interacts with stable radicals that possess approximately 658 times higher spin polarization. We utilized stable nitroxide radicals covalently attached along the hydrophobic tail of stearic acid molecules that incorporate themselves into surfactant-based micelle or vesicle structures. Here, we present a study of local water content and fluid viscosity inside oleate micelles and vesicles and Triton X-100 micelles to serve as model systems for soft molecular assemblies. This approach is unique because the amplification of the NMR signal is performed in bulk solution and under ambient conditions with site-specific spin labels that only detect the water that is directly interacting with the localized spin labels. Continuous wave (cw) electron spin resonance (ESR) analysis provides rotational dynamics of the spin-labeled molecular chain segments and local polarity parameters that can be related to hydration properties, whereas we show that DNP-enhanced (1)H NMR analysis of fluid samples directly provides translational water dynamics and permeability of the local environment probed by the spin label. Our technique therefore has the potential to become a powerful analysis tool, complementary to cw ESR, to study hydration characteristics of surfactant assemblies, lipid bilayers, or protein aggregates, where water dynamics is a key parameter of their structure and function. In this study, we find that there is significant penetration of water inside the oleate micelles with a higher average local water viscosity (approximately 1.8 cP) than in bulk water, and Triton X-100 micelles and oleate vesicle bilayers mostly exclude water while allowing for considerable surfactant chain motion and measurable water permeation through the soft structure.

Portable X-band system for solution state dynamic nuclear polarization.

This paper concerns instrumental approaches to obtain large dynamic nuclear polarization (DNP) enhancements in a completely portable system. We show that at fields of 0.35 T under ambient conditions and at X-band frequencies, 1H enhancements of >100-fold can be achieved using nitroxide radical systems, which is near the theoretical maximum for 1H polarization using the Overhauser effect at this field. These large enhancements were obtained using a custom built microwave transmitter and a commercial TE102 X-band resonant cavity. The custom built microwave transmitter is compact, so when combined with a permanent magnet it is readily transportable. Our commercial X-band resonator was modified to be tunable over a range of approximately 9.5-10 GHz, giving added versatility to our fixed field portable DNP system. In addition, a field adjustable Halbach permanent magnet has also been employed as another means for matching the electron spin resonance condition. Both portable setups provide large signal enhancements and with improvements in design and engineering, greater than 100-fold 1H enhancements are feasible.

A new model for Overhauser enhanced nuclear magnetic resonance using nitroxide radicals.

Nitroxide free radicals are the most commonly used source for dynamic nuclear polarization (DNP) enhanced nuclear magnetic resonance (NMR) experiments and are also exclusively employed as spin labels for electron spin resonance (ESR) spectroscopy of diamagnetic molecules and materials. Nitroxide free radicals have been shown to have strong dipolar coupling to (1)H in water, and thus result in large DNP enhancement of (1)H NMR signal via the well known Overhauser effect. The fundamental parameter in a DNP experiment is the coupling factor, since it ultimately determines the maximum NMR signal enhancements which can be achieved. Despite their widespread use, measurements of the coupling factor of nitroxide free radicals have been inconsistent, and current models have failed to successfully explain our experimental data. We found that the inconsistency in determining the coupling factor arises from not taking into account the characteristics of the ESR transitions, which are split into three (or two) lines due to the hyperfine coupling of the electron to the (14)N nuclei (or (15)N) of the nitric oxide radical. Both intermolecular Heisenberg spin exchange interactions as well as intramolecular nitrogen nuclear spin relaxation mix the three (or two) ESR transitions. However, neither effect has been taken into account in any experimental studies on utilizing or quantifying the Overhauser driven DNP effects. The expected effect of Heisenberg spin exchange on Overhauser enhancements has already been theoretically predicted and observed by Bates and Drozdoski [J. Chem. Phys. 67, 4038 (1977)]. Here, we present a new model for quantifying Overhauser enhancements through nitroxide free radicals that includes both effects on mixing the ESR hyperfine states. This model predicts the maximum saturation factor to be considerably higher by the effect of nitrogen nuclear spin relaxation. Because intramolecular nitrogen spin relaxation is independent of the nitroxide concentration, this effect is still significant at low radical concentrations where electron spin exchange is negligible. This implies that the only correct way to determine the coupling factor of nitroxide free radicals is to measure the maximum enhancement at different concentrations and extrapolate the results to infinite concentration. We verify our model with a series of DNP experimental studies on (1)H NMR signal enhancement of water by means of (14)N as well as (15)N isotope enriched nitroxide radicals.

Hyperpolarized water as an authentic magnetic resonance imaging contrast agent.

Pure water in a highly (1)H spin-polarized state is proposed as a contrast-agent-free contrast agent to visualize its macroscopic evolution in aqueous media by MRI. Remotely enhanced liquids for image contrast (RELIC) utilizes a (1)H signal of water that is enhanced outside the sample in continuous-flow mode and immediately delivered to the sample to obtain maximum contrast between entering and bulk fluids. Hyperpolarization suggests an ideal contrast mechanism to highlight the ubiquitous and specific function of water in physiology, biology, and materials because the physiological, chemical, and macroscopic function of water is not altered by the degree of magnetization. We present an approach that is capable of instantaneously enhancing the (1)H MRI signal by up to 2 orders of magnitude through the Overhauser effect under ambient conditions at 0.35 tesla by using highly spin-polarized unpaired electrons that are covalently immobilized onto a porous, water-saturated gel matrix. The continuous polarization of radical-free flowing water allowed us to distinctively visualize vortices in model reactors and dispersion patterns through porous media. A (1)H signal enhancement of water by a factor of -10 and -100 provides for an observation time of >4 and 7 s, respectively, upon its injection into fluids with a T(1) relaxation time of >1.5 s. The implications for chemical engineering or biomedical applications of using hyperpolarized solvents or physiological fluids to visualize mass transport and perfusion with high and authentic MRI contrast originating from water itself, and not from foreign contrast agents, are immediate.