Near Infrared Microspectroscopy, Fluorescence Microspectroscopy, Infrared Chemical Imaging and High-Resolution Nuclear Magnetic Resonance Analysis of Soybean Seeds, Embryos and Single Cells

Chemical analysis of soybean seeds, somatic embryos and single cells were carried out by Fourier Transform Infrared (FT-IR), Fourier Transform Near Infrared (FT-NIR) Microspectroscopy, Fluorescence and High-Resolution NMR (HR-NMR). The first FT-NIR chemical images of biological systems approaching 1 micron (1μ) resolution are presented here. Chemical images obtained by FT-NIR and FT-IR Microspectroscopy are presented for oil in soybean seeds and somatic embryos under physiological conditions. FT-NIR spectra of oil and proteins were obtained for volumes as small as 2μ3. Related, HR-NMR analyses of oil contents in somatic embryos are also presented here with nanoliter precision. Such 400 MHz 1H NMR analyses allowed the selection of mutagenized embryos with higher oil content (e.g. ~20%) compared to non-mutagenized control embryos. Moreover, developmental changes in single soybean seeds and/or somatic embryos may be monitored by FT-NIR with a precision approaching the picogram level. Indeed, detailed chemical analyses of oils and phytochemicals are now becoming possible by FT-NIR Chemical Imaging/ Microspectroscopy of single cells. The cost, speed and analytical requirements of plant breeding and genetic selection programs are fully satisfied by FT-NIR spectroscopy and Microspectroscopy for soybeans and soybean embryos. FT-NIR Microspectroscopy and Chemical Imaging are also shown to be potentially important in functional Genomics and Proteomics research through the rapid and accurate detection of high-content microarrays (HCMA). Multi-photon (MP), pulsed femtosecond laser NIR Fluorescence Excitation techniques were shown to be capable of Single Molecule Detection (SMD). Therefore, such powerful techniques allow for the most sensitive and reliable quantitative analyses to be carried out both in vitro and in vivo. Thus, MP NIR excitation for Fluorescence Correlation Spectroscopy (FCS) allows not only single molecule detection, but also molecular dynamics and high resolution, submicron imaging of femtoliter volumes inside living cells and tissues. These novel, ultra-sensitive and rapid NIR/FCS analyses have numerous applications in important research areas, such as: agricultural biotechnology, food safety, pharmacology, medical research and clinical diagnosis of viral diseases and cancers

Calibration of centre-of-mass energies at LEP 2 for a precise measurement of the W boson mass

The determination of the centre-of-mass energies for all LEP 2 running is presented. Accurate knowledge of these energies is of primary importance to set the absolute energy scale for the measurement of the W boson mass. The beam energy between 80 and 104 GeV is derived from continuous measurements of the magnetic bending field by 16 NMR probes situated in a number of the LEP dipoles. The relationship between the fields measured by the probes and the beam energy is defined in the NMR model, which is calibrated against precise measurements of the average beam energy between 41 and 61 GeV made using the resonant depolarisation technique. The validity of the NMR model is verified by three independent methods: the flux-loop, which is sensitive to the bending field of all the dipoles of LEP; the spectrometer, which determines the energy through measurements of the deflection of the beam in a magnet of known integrated field; and an analysis of the variation of the synchrotron tune with the total RF voltage. To obtain the centre-of-mass energies, corrections are then applied to account for sources of bending field external to the dipoles, and variations in the local beam energy at each interaction point. The relative error on the centre-of-mass energy determination for the majority of LEP 2 running is 1.2 x 10^{-4}, which is sufficiently precise so as not to introduce a dominant uncertainty on the W mass measurement.Comment: 79 pages, 45 figures, submitted to EPJ

Novel MRI Technologies for Structural and Functional Imaging of Tissues with Ultra-short T₂ Values

Conventional MRI has several limitations such as long scan durations, motion artifacts, very loud acoustic noise, signal loss due to short relaxation times, and RF induced heating of electrically conducting objects. The goals of this work are to evaluate and improve the state-of-the-art methods for MRI of tissue with short T₂, to prove the feasibility of in vivo Concurrent Excitation and Acquisition, and to introduce simultaneous electroglottography measurement during functional lung MRI

Emerging technologies for the non-invasive characterization of physical-mechanical properties of tablets

The density, porosity, breaking force, viscoelastic properties, and the presence or absence of any structural defects or irregularities are important physical-mechanical quality attributes of popular solid dosage forms like tablets. The irregularities associated with these attributes may influence the drug product functionality. Thus, an accurate and efficient characterization of these properties is critical for successful development and manufacturing of a robust tablets. These properties are mainly analyzed and monitored with traditional pharmacopeial and non-pharmacopeial methods. Such methods are associated with several challenges such as lack of spatial resolution, efficiency, or sample-sparing attributes. Recent advances in technology, design, instrumentation, and software have led to the emergence of newer techniques for non-invasive characterization of physical-mechanical properties of tablets. These techniques include near infrared spectroscopy, Raman spectroscopy, X-ray microtomography, nuclear magnetic resonance (NMR) imaging, terahertz pulsed imaging, laser-induced breakdown spectroscopy, and various acoustic- and thermal-based techniques. Such state-of-the-art techniques are currently applied at various stages of development and manufacturing of tablets at industrial scale. Each technique has specific advantages or challenges with respect to operational efficiency and cost, compared to traditional analytical methods. Currently, most of these techniques are used as secondary analytical tools to support the traditional methods in characterizing or monitoring tablet quality attributes. Therefore, further development in the instrumentation and software, and studies on the applications are necessary for their adoption in routine analysis and monitoring of tablet physical-mechanical properties

Tunable non-equilibrium dynamics: field quenches in spin ice

We present non-equilibrium physics in spin ice as a novel setting which combines kinematic constraints, emergent topological defects, and magnetic long range Coulomb interactions. In spin ice, magnetic frustration leads to highly degenerate yet locally constrained ground states. Together, they form a highly unusual magnetic state -- a "Coulomb phase" -- whose excitations are pointlike defects -- magnetic monopoles -- in the absence of which effectively no dynamics is possible. Hence, when they are sparse at low temperature, dynamics becomes very sluggish. When quenching the system from a monopole-rich to a monopole-poor state, a wealth of dynamical phenomena occur the exposition of which is the subject of this article. Most notably, we find reaction diffusion behaviour, slow dynamics due to kinematic constraints, as well as a regime corresponding to the deposition of interacting dimers on a honeycomb lattice. We also identify new potential avenues for detecting the magnetic monopoles in a regime of slow-moving monopoles. The interest in this model system is further enhanced by its large degree of tunability, and the ease of probing it in experiment: with varying magnetic fields at different temperatures, geometric properties -- including even the effective dimensionality of the system -- can be varied. By monitoring magnetisation, spin correlations or zero-field Nuclear Magnetic Resonance, the dynamical properties of the system can be extracted in considerable detail. This establishes spin ice as a laboratory of choice for the study of tunable, slow dynamics.Comment: (16 pages, 13 figures

The Application of Fast Correlation Methods with Dissolution DNP Enhanced NMR

Nuclear magnetic resonance (NMR) spectroscopy is a powerful analytical technique for chemistry and biochemistry. Since an inherent limitation of NMR is a lack of sensitivity, long experimental time with signal averaging or spins with high concentration are often required. An emerging technique, dissolution dynamic nuclear polarization (D-DNP), significantly enhances an NMR signal by several orders of magnitude. The large signal gains enable detection of spins with micromolar concentration and offer opportunities to various fields of applications. Two dimensional (2D) correlation spectroscopy plays an essential role in identifying molecular structure and dynamics. Because of the non-renewability of the hyperpolarized spin state, this property prevents the application of conventional 2D methods which rely on repetition of the experiment by successive increments of the indirect dimension. Therefore, it is important to find applicable methods to circumvent this problem. The present dissertation is focused on using fast single-scan 2D and pseudo-2D correlation methods to elucidate molecular structures and characterize physical parameters, such as diffusion and spin relaxation, with the goal of investigating reaction kinetics and mechanisms as well as studying membrane transport of metabolite. Since conventional 2D correlation spectroscopy is not compatible with D-DNP, an alternative way of collecting correlation information without obtaining an indirect spectral dimension is via off-resonance decoupling. Based on this concept, heteronuclear chemical shift correlations were determined in single scan DNP-enhanced NMR spectra under off resonance decoupling by Scaling of Heteronuclear Couplings by Optimal Tracking (SHOT) pulses, with the purpose of determining the identity of transient species and reaction mechanisms. Physical parameters of molecules, such as diffusion and spin relaxation, can further be characterized with single-scan correlation methods and used to examine membrane transport of metabolite. Ultrafast diffusion-Tv2 correlation Laplace NMR enables one to correlate spin relaxation and diffusion parameters in a single-scan. Diffusion-Tv2 correlation data was acquired by detecting hyperpolarized ^13C /^ 1H signals of small molecules, and maps were generated using inverse Laplace transform. The accurate determination of diffusion and Tv2 relaxation in homogeneous / inhomogeneous magnetic fields demonstrated the robustness of the method. The usability of hyperpolarized UF-LNMR is then demonstrated in the context of cell metabolism, by investigating the conversion of pyruvate to lactate in the cultures of mouse 4T1 cancer cells. We show that ^13C ultrafast diffusion-Tv2 relaxation correlation measurements, with the sensitivity enhanced by several orders of magnitude by D-DNP, allows the determination of the extra- vs. intracellular location of metabolites in the cells due to their significantly different values of diffusion coefficients and Tv2 relaxation time

Muon spin spectroscopy: magnetism, soft matter and the bridge between the two

LS would like to acknowledge financial support from the Swiss National Science Foundation, grant numbers PBFRP2-138632 and PBFRP2-142820. AD would like to acknowledge financial support from the UK Engineering and Physical Sciences Research Council, grant number EP/G054568/1, the European Union Seventh Framework Programme project NMP3-SL- 2011-263104 ‘HINTS’ and the European Research Council project ‘Muon Spin Spectroscopy of Excited States (MuSES)’ proposal number 307593LS would like to acknowledge financial support from the Swiss National Science Foundation, grant numbers PBFRP2-138632 and PBFRP2-142820. AD would like to acknowledge financial support from the UK Engineering and Physical Sciences Research Council, grant number EP/G054568/1, the European Union Seventh Framework Programme project NMP3-SL- 2011-263104 ‘HINTS’ and the European Research Council project ‘Muon Spin Spectroscopy of Excited States (MuSES)’ proposal number 307593LS would like to acknowledge financial support from the Swiss National Science Foundation, grant numbers PBFRP2-138632 and PBFRP2-142820. AD would like to acknowledge financial support from the UK Engineering and Physical Sciences Research Council, grant number EP/G054568/1, the European Union Seventh Framework Programme project NMP3-SL- 2011-263104 ‘HINTS’ and the European Research Council project ‘Muon Spin Spectroscopy of Excited States (MuSES)’ proposal number 30759

Doctor of Philosophy

dissertationMinimally invasive thermal therapy under Magnetic Resonance Imaging (MRI) guidance is becoming popular with several applications in the process of getting FDA approval. The ability to determine in near real-time the temperature map of a tumor and its surrounding tissue makes MR thermometry very attractive and well suited for thermal treatment. The proton resonance frequency shift (PRF) is currently the gold standard method for temperature monitoring using MRI. However, its incapacity to measure temperature in fatty tissue limits the scope of its applicability. The spin lattice relaxation time T1, on the other hand, has shown good temperature sensitivity and works well in all types of tissues. In this dissertation, we have addressed a number of challenges currently affecting MRI thermometry. A non-CPMG Turbo Spin Echo (TSE) sequence has been implemented to monitor the temperature rise due to the high RF power deposition inherent to this sequence at high field (3T and higher). This new implementation allows TSE sequences to be used safely without altering their high contrast properties which make them appealing in clinical settings. Tissue damage assessment during thermal therapy is critical for the safety of the patient. We have developed a new hybrid PRF-T1 sequence that has the capability to provide simultaneously in near real-time the temperature map and T1 information, which is a good indication of the state of the tissue. The simplicity and the real-time capability of the newly developed sequence make it an ideal tool for tissue damage assessment. Temperature monitoring during thermal therapy in organs with large fat content have been hindered by the lack of an MRI thermometry method that can provide simultaneous temperature in fat and aqueous tissue. A new sequence and acquisition scheme have been developed to address this issue. In sum, this dissertation proposed several pulse sequence implementation techniques and an acquisition scheme to overcome some of the limitations of MR thermometry