Chemical Shift Imaging Experiments Research Articles

Measurement of the pKa Values of Organic Molecules in Aqueous-Organic Solvent Mixtures by 1H NMR without External Calibrants.

Aqueous-organic solvent mixtures are commonly used for reactions or analyses, where the components of a system are insoluble in pure water. The acid dissociation constant is an important property to measure in these media as it determines the charge state, solubility, and reactivity of a molecule. While NMR spectroscopy is an established tool for the measurement of pKa in water, its use in aqueous-organic solvents is greatly hindered by the requirement for external calibrants on which a working pH scale is set. Such calibrants include buffer solutions, "anchor" molecules with known pKa values, and pH electrodes that have undergone lengthy calibration procedures in the solvent mixture of interest. However, such calibrations are often inconvenient to perform, while literature pKa data covering the required range may not be available at the solvent composition or the temperature of interest. Here, we present a method to determine pKa in aqueous-organic solvents directly by NMR. We first determine pKa of an organic acid such as 2,6-dihydroxybenzoic acid (2,6-DHB) by measuring its 1H chemical shift as a function of concentration along a concentration gradient using chemical shift imaging (CSI). Using 2,6-DHB as a reference, we then determine pKa of less acidic molecules in single CSI experiments via the variation of their 1H chemical shifts along pH gradients. As proof of concept, we determine the pKa values of organic acids and bases up to pKa 10 in 50% (v/v) 1-propanol/water, 50% (v/v) dimethyl sulfoxide/water, and 30% (v/v) acetonitrile/water and obtain good agreement with the literature values.

Deuterium MRSI characterizations of glucose metabolism in orthotopic pancreatic cancer mouse models.

Detecting and mapping metabolism in tissues represents a major step in detecting, characterizing, treating and understanding cancers. Recently introduced deuterium metabolic imaging techniques could offer a noninvasive route for the metabolic imaging of animals and humans, based on using 2 H magnetic resonance spectroscopic imaging (MRSI) to detect the uptake of deuterated glucose and the fate of its metabolic products. In this study, 2 H6,6' -glucose was administered to mice cohorts that had been orthotopically implanted with two different models of pancreatic ductal adenocarcinoma (PDAC), involving PAN-02 and KPC cell lines. As the tumors grew, 2 H6,6' -glucose was administered as bolii into the animals' tail veins, and 2 H MRSI images were recorded at 15.2T. 2D phase-encoded chemical shift imaging experiments could detect a signal from this deuterated glucose immediately after the bolus injection for both the PDAC models, reaching a maximum in the animals' tumors ~ 20 min following administration, and nearly total decay after ~ 40 min. The main metabolic reporter of the cancers was the 2 H3,3' -lactate signal, which MRSI could detect and localize on the tumors when these were 5mm or more in diameter. Lactate production time traces varied slightly with the animal and tumor model, but in general lactate peaked at times of 60 min or longer following injection, reaching concentrations that were ~ 10-fold lower than those of the initial glucose injection. This 2 H3,3' -lactate signal was only visible inside the tumors. 2 H-water could also be detected as deuterated glucose's metabolic product, increasing throughout the entire time course of the experiment from its ≈10 mM natural abundance background. This water resonance could be imaged throughout the entire abdomen of the animals, including an enhanced presence in the tumor, but also in other organs like the kidney and bladder. These results suggest that deuterium MRSI may serve as a robust, minimally invasive tool for the monitoring of metabolic activity in pancreatic tumors, capable of undergoing clinical translation and supporting decisions concerning treatment strategies. Comparisons with in vivo metabolic MRI experiments that have been carried out in other animal models are presented and their differences/similarities are discussed.

Multinuclear absolute magnetic resonance thermometry

Non-invasive measurement of absolute temperature is important for proper characterization of various pathologies and for evaluation of thermal dose during interventional procedures. The proton (hydrogen nucleus) magnetic resonance (MR) frequency shift method can be used to map relative temperature changes. However, spatiotemporal variations in the main magnetic field and the lack of local internal frequency reference challenge the determination of absolute temperature. Here, we introduce a multinuclear method for absolute MR thermometry, based on the fact that the hydrogen and sodium nuclei exhibit a unique and distinct characteristic frequency dependence with temperature and with electrolyte concentration. A one-to-one mapping between the precession frequency difference of the two nuclei and absolute temperature is demonstrated. Proof-of-concept experiments were conducted in aqueous solutions with different NaCl concentrations, in agarose gel samples, and in freshly excised ex vivo mouse tissues. One-dimensional chemical shift imaging experiments also demonstrated excellent agreement with infrared measurements.

Dynamic Imaging of Glucose and Lactate Metabolism by 13C-MRS without Hyperpolarization

Metabolic reprogramming is one of the defining features of cancer and abnormal metabolism is associated with many other pathologies. Molecular imaging techniques capable of detecting such changes have become essential for cancer diagnosis, treatment planning, and surveillance. In particular, 18F-FDG (fluorodeoxyglucose) PET has emerged as an essential imaging modality for cancer because of its unique ability to detect a disturbed molecular pathway through measurements of glucose uptake. However, FDG-PET has limitations that restrict its usefulness in certain situations and the information gained is limited to glucose uptake only.13C magnetic resonance spectroscopy theoretically has certain advantages over FDG-PET, but its inherent low sensitivity has restricted its use mostly to single voxel measurements unless dissolution dynamic nuclear polarization (dDNP) is used to increase the signal, which brings additional complications for clinical use. We show here a new method of imaging glucose metabolism in vivo by MRI chemical shift imaging (CSI) experiments that relies on a simple, but robust and efficient, post-processing procedure by the higher dimensional analog of singular value decomposition, tensor decomposition. Using this procedure, we achieve an order of magnitude increase in signal to noise in both dDNP and non-hyperpolarized non-localized experiments without sacrificing accuracy. In CSI experiments an approximately 30-fold increase was observed, enough that the glucose to lactate conversion indicative of the Warburg effect can be imaged without hyper-polarization with a time resolution of 12s and an overall spatial resolution that compares favorably to 18F-FDG PET.

ViP-CSI: Virtual Phantom Chemical Shift Imaging

To apply the Virtual Phantom (ViP) method for generating reference signals, to the chemical shift imaging (CSI) technique. The ViP method, previously used for generating reference images in magnetic resonance imaging (MRI), was here extended to generate reference peaks in the MR spectra acquired with a 2D-CSI sequence. Theoretical analysis was carried out to design the ViP signal. ViP-2D-CSI experiments were performed on a 4.7 Tesla MR scanner. Data analysis was carried out with the jMRUI software to fit the ViP peaks to determine their amplitude and frequency. Based on the theoretical analysis, it was found that the ViP peak could be generated by transmitting the ViP signal only in one repetition time of the 2D-CSI sequence. The amplitude and frequency of the ViP peak could be precisely adjusted and fine-tuned. The ViP peak was uniform over all CSI voxels, both in amplitude and in frequency. Furthermore, a good stability of the ViP reference amplitude and frequency was observed. The ViP method provides a means to generate a reference MR peak in 2D-CSI experiments. This could be of interest for signal quantification in CSI experiments.

Fast Padé Transform Accelerated CSI for Hyperpolarized MRS.

The fast Padé transform (FPT) is a method of spectral analysis that can be used to reconstruct nuclear magnetic resonance spectra from truncated free induction decay signals with superior robustness and spectral resolution compared with conventional Fourier analysis. The aim of this study is to show the utility of FPT in reducing of the scan time required for hyperpolarized 13C chemical shift imaging (CSI) without sacrificing the ability to resolve a full spectrum. Simulations, phantom, and in vivo hyperpolarized [1-13C] pyruvate CSI data were processed with FPT and compared with conventional analysis methods. FPT shows improved stability and spectral resolution on truncated data compared with the fast Fourier transform and shows results that are comparable to those of the model-based fitting methods, enabling a reduction in the needed acquisition time in 13C CSI experiments. Using FPT can reduce the readout length in the spectral dimension by 2-6 times in 13C CSI compared with conventional Fourier analysis without sacrificing the spectral resolution. This increased speed is crucial for 13C CSI because T1 relaxation considerably limits the available scan time. In addition, FPT can also yield direct quantification of metabolite concentration without the additional peak analysis required in conventional Fourier analysis.

Heteronuclear Micro-Helmholtz Coil Facilitates µm-Range Spatial and Sub-Hz Spectral Resolution NMR of nL-Volume Samples on Customisable Microfluidic Chips.

We present a completely revised generation of a modular micro-NMR detector, featuring an active sample volume of ∼ 100 nL, and an improvement of 87% in probe efficiency. The detector is capable of rapidly screening different samples using exchangeable, application-specific, MEMS-fabricated, microfluidic sample containers. In contrast to our previous design, the sample holder chips can be simply sealed with adhesive tape, with excellent adhesion due to the smooth surfaces surrounding the fluidic ports, and so withstand pressures of ∼2.5 bar, while simultaneously enabling high spectral resolution up to 0.62 Hz for H2O, due to its optimised geometry. We have additionally reworked the coil design and fabrication processes, replacing liquid photoresists by dry film stock, whose final thickness does not depend on accurate volume dispensing or precise levelling during curing. We further introduced mechanical alignment structures to avoid time-intensive optical alignment of the chip stacks during assembly, while we exchanged the laser-cut, PMMA spacers by diced glass spacers, which are not susceptible to melting during cutting. Doing so led to an overall simplification of the entire fabrication chain, while simultaneously increasing the yield, due to an improved uniformity of thickness of the individual layers, and in addition, due to more accurate vertical positioning of the wirebonded coils, now delimited by a post base plateau. We demonstrate the capability of the design by acquiring a 1H spectrum of ∼ 11 nmol sucrose dissolved in D2O, where we achieved a linewidth of 1.25 Hz for the TSP reference peak. Chemical shift imaging experiments were further recorded from voxel volumes of only ∼ 1.5nL, which corresponded to amounts of just 1.5 nmol per voxel for a 1 M concentration. To extend the micro-detector to other nuclei of interest, we have implemented a trap circuit, enabling heteronuclear spectroscopy, demonstrated by two 1H/13C 2D HSQC experiments.

Quantitative mapping of chemical compositions with MRI using compressed sensing

In this work, a magnetic resonance (MR) imaging method for accelerating the acquisition time of two dimensional concentration maps of different chemical species in mixtures by the use of compressed sensing (CS) is presented. Whilst 2D-concentration maps with a high spatial resolution are prohibitively time-consuming to acquire using full k-space sampling techniques, CS enables the reconstruction of quantitative concentration maps from sub-sampled k-space data. First, the method was tested by reconstructing simulated data. Then, the CS algorithm was used to reconstruct concentration maps of binary mixtures of 1,4-dioxane and cyclooctane in different samples with a field-of-view of 22mm and a spatial resolution of 344μm×344μm. Spiral based trajectories were used as sampling schemes. For the data acquisition, eight scans with slightly different trajectories were applied resulting in a total acquisition time of about 8min. In contrast, a conventional chemical shift imaging experiment at the same resolution would require about 17h. To get quantitative results, a careful weighting of the regularisation parameter (via the L-curve approach) or contrast-enhancing Bregman iterations are applied for the reconstruction of the concentration maps. Both approaches yield relative errors of the concentration map of less than 2mol-% without any calibration prior to the measurement. The accuracy of the reconstructed concentration maps deteriorates when the reconstruction model is biased by systematic errors such as large inhomogeneities in the static magnetic field. The presented method is a powerful tool for the fast acquisition of concentration maps that can provide valuable information for the investigation of many phenomena in chemical engineering applications.

A subspace approach to high‐resolution spectroscopic imaging

To accelerate spectroscopic imaging using sparse sampling of (k,t)-space and subspace (or low-rank) modeling to enable high-resolution metabolic imaging with good signal-to-noise ratio. The proposed method, called SPectroscopic Imaging by exploiting spatiospectral CorrElation, exploits a unique property known as partial separability of spectroscopic signals. This property indicates that high-dimensional spectroscopic signals reside in a very low-dimensional subspace and enables special data acquisition and image reconstruction strategies to be used to obtain high-resolution spatiospectral distributions with good signal-to-noise ratio. More specifically, a hybrid chemical shift imaging/echo-planar spectroscopic imaging pulse sequence is proposed for sparse sampling of (k,t)-space, and a low-rank model-based algorithm is proposed for subspace estimation and image reconstruction from sparse data with the capability to incorporate prior information and field inhomogeneity correction. The performance of the proposed method has been evaluated using both computer simulations and phantom studies, which produced very encouraging results. For two-dimensional spectroscopic imaging experiments on a metabolite phantom, a factor of 10 acceleration was achieved with a minimal loss in signal-to-noise ratio compared to the long chemical shift imaging experiments and with a significant gain in signal-to-noise ratio compared to the accelerated echo-planar spectroscopic imaging experiments. The proposed method, SPectroscopic Imaging by exploiting spatiospectral CorrElation, is able to significantly accelerate spectroscopic imaging experiments, making high-resolution metabolic imaging possible.

In vivo three‐dimensional molecular imaging with Biosensor Imaging of Redundant Deviation in Shifts (BIRDS) at high spatiotemporal resolution

Spectroscopic signals which emanate from complexes between paramagnetic lanthanide (III) ions (e.g. Tm(3+)) and macrocyclic chelates (e.g. 1,4,7,10-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate, or DOTMA(4-)) are sensitive to physiology (e.g. temperature). Because nonexchanging protons from these lanthanide-based macrocyclic agents have relaxation times on the order of a few milliseconds, rapid data acquisition is possible with chemical shift imaging (CSI). Thus, Biosensor Imaging of Redundant Deviation in Shifts (BIRDS) which originate from nonexchanging protons of these paramagnetic agents, but exclude water proton detection, can allow molecular imaging. Previous two-dimensional CSI experiments with such lanthanide-based macrocyclics allowed acquisition from ~12-μL voxels in rat brain within 5 min using rectangular encoding of k space. Because cubical encoding of k space in three dimensions for whole-brain coverage increases the CSI acquisition time to several tens of minutes or more, a faster CSI technique is required for BIRDS to be of practical use. Here, we demonstrate a CSI acquisition method to improve three-dimensional molecular imaging capabilities with lanthanide-based macrocyclics. Using TmDOTMA(-), we show datasets from a 20 × 20 × 20-mm(3) field of view with voxels of ~1 μL effective volume acquired within 5 min (at 11.7 T) for temperature mapping. By employing reduced spherical encoding with Gaussian weighting (RESEGAW) instead of cubical encoding of k space, a significant increase in CSI signal is obtained. In vitro and in vivo three-dimensional CSI data with TmDOTMA(-), and presumably similar lanthanide-based macrocyclics, suggest that acquisition using RESEGAW can be used for high spatiotemporal resolution molecular mapping with BIRDS.

Localization errors in MR spectroscopic imaging due to the drift of the main magnetic field and their correction

To analyze the effect of B0 field drift on multivoxel MR spectroscopic imaging and to propose an approach for its correction. It is shown, both theoretically and in a phantom, that for ∼30 min acquisitions a linear B0 drift (∼0.1 ppm/h) will cause localization errors that can reach several voxels (centimeters) in the slower varying phase encoding directions. An efficient and unbiased estimator is proposed for tracking the drift by interleaving short (∼ T2*), nonlocalized acquisitions on the nonsuppressed water each pulse repetition time, as shown in 10 volunteers at 1.5 and 3 T. The drift is shown to be predominantly linear in both the phantom and volunteers at both fields. The localization errors are observed and quantified in both phantom and volunteers. The unbiased estimator is shown to reliably track the instantaneous frequency in vivo despite only using a small portion of the FID. Contrary to single-voxel MR spectroscopy, where it leads to line broadening, field drift can lead to localization errors in the longer chemical shift imaging experiments. Fortunately, this drift can be obtained at a negligible cost to sequence timing, and corrected for in post processing.

Application of compressed sensing to in vivo 3D 19F CSI

This study shows how applying compressed sensing (CS) to 19F chemical shift imaging (CSI) makes highly accurate and reproducible reconstructions from undersampled datasets possible. The missing background signal in 19F CSI provides the required sparsity needed for application of CS. Simulations were performed to test the influence of different CS-related parameters on reconstruction quality. To test the proposed method on a realistic signal distribution, the simulation results were validated by ex vivo experiments. Additionally, undersampled in vivo 3D CSI mouse datasets were successfully reconstructed using CS. The study results suggest that CS can be used to accurately and reproducibly reconstruct undersampled 19F spectroscopic datasets. Thus, the scanning time of in vivo 19F CSI experiments can be significantly reduced while preserving the ability to distinguish between different 19F markers. The gain in scan time provides high flexibility in adjusting measurement parameters. These features make this technique a useful tool for multiple biological and medical applications.

Brain temperature and pH measured by 1H chemical shift imaging of a thulium agent

Temperature and pH are two of the most important physiological parameters and are believed to be tightly regulated because they are intricately related to energy metabolism in living organisms. Temperature and/or pH data in mammalian brain are scarce, however, mainly because of lack of precise and non-invasive methods. At 11.7 T, we demonstrate that a thulium-based macrocyclic complex infused through the bloodstream can be used to obtain temperature and pH maps of rat brain in vivo by (1)H chemical shift imaging (CSI) of the sensor itself in conjunction with a multi-parametric model that depends on several proton resonances of the sensor. Accuracies of temperature and pH determination with the thulium sensor - which has a predominantly extracellular presence - depend on stable signals during the course of the CSI experiment as well as redundancy for temperature and pH sensitivities contained within the observed signals. The thulium-based method compared well with other methods for temperature ((1)H MRS of N-acetylaspartate and water; copper-constantan thermocouple wire) and pH ((31)P MRS of inorganic phosphate and phosphocreatine) assessment, as established by in vitro and in vivo studies. In vitro studies in phantoms with two compartments of different pH value observed under different ambient temperature conditions generated precise temperature and pH distribution maps. In vivo studies in alpha-chloralose-anesthetized and renal-ligated rats revealed temperature (33-34 degrees C) and pH (7.3-7.4) distributions in the cerebral cortex that are in agreement with observations by other methods. These results show that the thulium sensor can be used to measure temperature and pH distributions in rat brain in vivo simultaneously and accurately using Biosensor Imaging of Redundant Deviation in Shifts (BIRDS).

NMR Microscopy of an “Anisotropic” Polymer

AbstractTwo different samples of high‐density polyethylene (HDPE) have been studied. One (isotropic) is extracted from the material core whereas the other (anisotropic) involves two sides which have been in contact with the injection mold. It is observed by NMR microscopy (using radiofrequency field gradients) that these two sides favor toluene penetration into the material. The distribution of toluene nuclear spin relaxation times (extracted from proton T1 and T2 images) exhibits likewise important differences between the two samples. These differences can be accounted for by partial molecular ordering at the vicinity of the “mold sides”. Finally, in investigating the anisotropic sample (without solvent), three different phases (two amorphous and one crystalline) are revealed by 13C chemical shift imaging experiments (performed with radiofrequency field gradients under CP/MAS conditions). Each amorphous component is preferentially present at one of the two “mold sides”.

Accurate phosphorus metabolite images of the human heart by 3D acquisition-weighted CSI.

Fourier imaging modalities suffer from significant signal contamination between adjacent voxels, especially when the spatial resolution is comparable to the size of the anatomical structures. This contamination can be positive or negative, depending on the spatial response function and the geometry of the object. Such a situation arises in human cardiac (31)P chemical shift imaging (CSI). Acquisition-weighted CSI reduces this contamination substantially, which is demonstrated by comparing conventional CSI to Hanning-weighted 3D (31)P-CSI experiments in 13 healthy volunteers at 2 T. The nominal spatial resolution and the total number of scans were identical for both experiments. The improved spatial response function of the acquisition-weighted experiment led to a significantly (P < 0.0001) higher myocardial PCr/ATP ratio (2.05 +/- 0.31, mean +/- SD, N = 33, corrected for saturation and blood contribution) compared to the conventional CSI experiment (1.60 +/- 0.46). This is explained by the absence of negative contamination from skeletal muscle, which also resulted in an increase of the observed SNR (from 5.4 +/- 1.4 to 7.2 +/- 1.4 for ATP). With acquisition-weighted CSI, metabolic images with a nominal resolution of 16 ml could be obtained in a measurement time of 30 min. After correction for the inhomogeneous B(1) field of the surface coil, these images show uniform ATP distribution in the entire myocardium, including the posterior wall.

A quantitative assessment of liver metabolites during jaundice using three dimensional phosphorus chemical shift imaging☆

Phosphorus metabolites in the jaundiced rat liver were studied by three-dimensional phosphorus chemical shift imaging (CSI). Animals were studied at 1, 2, and 3 weeks post-ligation of the common bile duct. Quantitation of metabolites was performed using an external standard. Metabolite T 1 values were assessed in CSI experiments on normal untreated animals. High-performance liquid chromatography (HPLC) was used to measure adenine nucleotides in a separate group of jaundiced rats. 3D-CSI did not detect significant changes in NTP in jaundiced animals relative to baseline controls. At two and three weeks post bile duct ligation, pH was significantly elevated. HPLC data comparing ATP levels to baseline controls also detected no change except for elevated ATP detected on Day 21. 31P NMR chemical shift imaging may be used to assess liver metabolites under conditions of stress such as jaundice. However, absolute quantitation requires careful attention to many factors including point spread function, correct T 1 values, and adequate signal-to-noise ratio.

Beyond k-Space: Spectral Localization Using Higher Order Gradients

Chemical shift imaging (CSI) often suffers from the inconvenient shape of its spatial response function (SRF), which affects both localization and signal-to-noise ratio. Replacing the magnetic field gradients for phase encoding by higher order magnetic fields allows a better adjustment of the SRF to the structures in the sample. We combined this principle with the SLOOP (spectral localization with optimal pointspread function) technique to simultaneously obtain spectra from several arbitrarily shaped compartments within a sample. Linear combinations of the fields of the shim coils are used to generate the pulsed fields for phase encoding. Their shapes are matched to the given sample geometry by numerical optimization. Using this method, spectra from a phantom were obtained that show a higher signal-to-noise ratio and a strongly reduced contamination compared to an equivalent CSI experiment.

Measurement of Convection and Temperature Profiles in Liquid Samples

This paper describes a method for measuring the rate of convective flow in a liquid sample used for high-resolution NMR. The measurement is straightforward and achieves a clean separation of convection from other effects such as diffusion and relaxation. Convection results from temperature gradients within the sample, and it is shown how these can be measured with the aid of a simple chemical shift imaging experiment of a sample whose spectrum shows a strong and well characterized temperature dependence. The use of these two methods is illustrated by showing how the rate of convection and the temperature profile depend on the solvent, temperature, and gas flow rate of the temperature regulating system. It is also shown that broadband 13C decoupling results in significant temperature gradients and associated convection.

Spatial distribution of deoxymyoglobin in human muscle: an index of local tissue oxygenation.

The proximal histidyl NdeltaH signal of myoglobin is detectable in 1H NMR spectra of myocardial and skeletal muscle, and its intensity reflects the intracellular oxygenation. At 1.5 Tesla (T), the typical field strength of clinical magnetic resonance imaging (MRI) magnets, the paramagnetic relaxation contribution decreases sufficiently to permit the implementation of chemical shift imaging technique to map the spatial distribution of the deoxy Mb NdeltaH signal from human gastrocnemius muscle. One and two-dimensional chemical shift imaging experiments reveal clearly the localized deoxy Mb signal in muscle and consequently the spatial distribution of the cellular oxygenation. The results indicate the feasibility to assess the pO2 in tissue regions and to directly study the regulation of oxidative metabolism in human tissue.

Theoretical Evaluation and Comparison of Fast Chemical Shift Imaging Methods

Magnetic resonance chemical shift imaging (CSI) is becoming the method of choice for localized NMR spectroscopic examinations, allowing simultaneous detection of NMR spectra from a large number of voxels. The main limitation of these methods is their long experimental duration. A number of fast CSI experiments have been presented, promising to reduce that duration. In this contribution the criteria for evaluating and optimizing the sensitivity of fast CSI experiments are elaborated. For a typical experiment in the human brain, the performance of various methods is compared. While conventional CSI provides optimal sensitivity per unit time, it is shown in which circumstances fast sequences allow a shorter experimental duration. Using these results, the best method for any experimental requirements can be selected.