APPLICATION STUDY ON SOLVENT SIGNAL SUPPRESSION WITH SELECTIVE EXCITATION TECHNIQUE IN COMBINATION WITH PFG

对结合脉冲梯度场的选择激发技术在溶剂峰抑制中的四种可行序列进行比较,实验表明非对称的双梯度自旋回波序列在谱图的相位特性和溶剂峰抑制效果两方面均是最佳的方案,并将其推广应用到溶剂峰抑制的gCOSY实验中,获得较理想的溶剂峰抑制效果.     

Determination and assignment of heteronuclear long-range coupling constants. Methods based on semiselective INEPT

One-dimensional methods for the determination and assignment of heteronuclear ¹H-X (X = rare spin-z) coupling constants based on the semiselective polarization transfer via INEPT pulse sequence are proposed. Here the selectivity of the polarization transfer plays a positive role with respect to the sensitivity of the measurement and purity of the observed multiplets. In nonrefocused experiments the acquired antiphase multiplets enable an unambiguous assignment of long-range couplings of a preselected proton. The analysis of such multiplets is also discussed. In the refocused version the purging pulse (INEPT+) was used to provide pure in-phase multiplets. The spectral editing technique DISCO was applied to simplify the spectra and to extract the couplings from complex multiplets. Finally, the modified INEPT experiments which combine semiselective polarization transfer with selective proton decoupling are proposed.     

The dipolar and exchange contributions to the spin-lattice relaxation of the imino protons in poly(rA)· poly(rU)

Selective, semiselective, and biselective excitation has been used to study the spin-lattice relaxation of the hydrogen-bonded imino protons in the sonicated, double-stranded RNA polymer poly(rA)· poly(rU). The spin-lattice relaxation of the imino protons has contributions from both dipolar interactions and exchange with the solvent, and the relative contribution of each to the observed rate depends on temperature. When exchange is slow compared to the dipolar contribution, the two components can be resolved by measuring the relaxation rate with and without prior inversion of the solvent peak. When exchange is fast, the contributions can be resolved by comparing the initial selective relaxation rate with the rate of saturation transfer when the solvent peak is inverted. This allows the exchange and dipolar contributions to be separated at temperatures well below the duplex melting temperatures and provides an easy way to study the mechanism of proton exchange with the solvent.     

1D Determination of Long-Range Heteronuclear Coupling Constants by Gradient Enhanced SIMBA

Two¹H-detected one-dimensional gradient enhanced experiments and corresponding reference experiments are proposed for the measurement of heteronuclear long-rangeⁿJ_CHcoupling constants in organic molecules. These experiments use inverse detection and selective carbon excitation, provide excellent suppression of protons not coupled to¹³C, and are able to measure couplings from regions that are crowded in both the proton and carbon frequency ranges. The experiments are applied to determine the long-rangeⁿJ_CHcouplings in the sucrose molecule, including coupling across the glycosidic linkage. These pulse sequences are shown to provide accurate and efficient measurements of coupling constants that would be difficult or time-consuming to measure by other techniques.     

Measurement of transverse relaxation in coupled spin systems by semi-selective refocusing: Evidence for interference of CSA and dipolar relaxation

A novel spin-echo experiment is proposed that uses semiselective Gaussian pulse cascades for refocusing in systems with homonuclear couplings. The experiment allows one to obtain evidence of differential T₂ effects due to cross-correlation of proton chemical shift anisotropy and proton-proton dipolar interactions in a small aromatic compound at 9.4 T. The spin-echo experiment is also applied to measure transverse relaxation rates for selected multiplets of the cyclic undecapeptide cyclosporin. Possible extensions to multiple-quantum spin echo-spectroscopy are indicated.     

RF Gradient BIRD/TANGO Sequence to Eliminate Uncoupled Magnetization

A modification of the BIRD and TANGO sequences is presented which employs radiofrequency field gradients to eliminate the net magnetization from uncoupled spins, while completely preserving coupled magnetization. The standard BIRD and TANGO sequences cause selective nutation of protons directly bound to a coupling partner, while returning uncoupled magnetization to +z. These sequences lend themselves naturally to modification using RF gradients, which require no increase in pulse-sequence complexity while providing substantial suppression of uncoupled resonances and elimination of typical antiphase and multiple-quantum error terms that arise from improperly set pulse lengths or delays. In the RF-gradient BIRD/TANGO sequence, the uncoupled magnetization is dephased in a plane orthogonal to the RF axis, while the desired signal components are refocused, effectively in a rotary echo. The sequence has applications to solvent suppression and selective isotopomer excitation. It is demonstrated for selective excitation of the satellites in a sample of chloroform, yielding suppression of the uncoupled magnetization by a factor of approximately 800.     

Absolute signs of hyperfine coupling constants as determined by pulse ENDOR of polarized radical pairs

A novel method that allows the determination of absolute signs of hyperfine coupling constants in polarized radical pair (RP) pulse electron-nuclear double resonance (ENDOR) spectra is presented, The variable mixing time (VMT) ENDOR method used here leads to a separation of ENDOR transitions originating from different electron spin manifolds by employing their dependence on the time-dependent parameters of the pulse sequence. The simple kinetic model of the RP VMT ENDOR experiment shows very good agreement with the experiments performed on the P ₇₀₀ ^.+ A ₁ ^.- RP in photosystem I. This method relies on the selective excitation of absorptive or emissive lines of one radical in the RP EPR spectrum and therefore requires high spectral resolution. This condition was fulfilled for the system studied at the low-field edge of the RP EPR spectrum obtained at Q-band. The method presented here has a very high sensitivity and does not require any equipment additional to the one used for RP pulse ENDOR. The VMT ENDOR method offers the possibility for selective suppression of signals from different electron spin manifolds.     

MUSIC and Aromatic Residues: Amino Acid Type-Selective H–N Correlations, III

Amino acid type-selective experiments can help to remove ambiguities in automated assignment procedures for ¹⁵N/¹³C-labeled proteins. Here we present five triple-resonance experiments that yield amino acid type-selective ¹H–¹⁵N correlations for aromatic amino acids. Four of the novel experiments are based on the MUSIC coherence transfer scheme that replaces the initial INEPT transfer and is selective for CH₂. The MUSIC sequence is combined with selective excitation pulses to create experiments for Trp (W-HSQC) as well as Phe, Tyr, and His (FYH-HSQC). In addition, an experiment selective for Trp H¹–N¹ is presented. The new experiments are recorded as two-dimensional experiments and their performance is demonstrated with the application to a protein domain of 115 amino acids.     

Selective excitation in Fourier transform nuclear magnetic resonance. 1978

The applications of frequency-selective excitation methods in Fourier transform NMR are discussed, and a simple technique is described for selective excitation of a narrow frequency region of a high-resolution NMR spectrum in a Fourier transform spectrometer. A regular sequence of identical radiofrequency pulses of small flip angle exerts a strong cumulative effect on magnetizations close to resonance with the transmitter frequency or one of a set of equally spaced sidebands separated by the pulse repetition rate. All other magnetizations precess through an incomplete number of full rotations between pulses, and are caught by successive pulses at an ever changing phase of their precession, which destroys the cumulative effect. The motion of the various nuclear magnetization vectors may be described pictorially according to the Bloch equations, neglecting relaxation during the pulse sequence. A general theory is presented for selective or “tailored” excitation by an arbitrary modulation of the radiofrequency transmitter signal. It confirms earlier conclusions that the frequency-domain excitation spectrum corresponds to the Fourier transform of the transmitter modulation pattern, provided that the NMR response remains linear. The excitation spectra calculated for the selective pulse sequence by these two alternative approaches show good agreement within their respective limitations. A number of practical applications of selective excitation are explored, including solvent peak suppression, the detection of partial spectra from individual chemical sites, selective studies of relaxation and slow chemical exchange, and holeburning or localized saturation.     

Projection Presaturation. III. Accurate Selective Excitation or Presaturation of the Regions of Tailored Shape in the Presence of Short-T1 Species

A new method is described which further increases the accuracy of localization by projection presaturation (PP), a technique for multidimensional spatial localization of contoured regions of interest (ROIs), whose shape, size, position, and number can be chosen arbitrarily (J. Magn. Reson.90, 313, 1990). By applying a nonselective radiofrequency inversion pulse after the PP saturation cycle and nulling the residual longitudinal magnetization of the region outside the ROI, the outer-volume signal is suppressed to the "noise" level, thus improving the accuracy of the basic PP localization, even when saturating a large exterior volume of short-T₁ species. The ability of the method to perform spatial and spectral localization simultaneously is also described. Alternatively, the single-shot methods for selective saturation (instead of selective excitation) of single or multiple contoured regions at arbitrary positions in an extended volume are presented. In particular, the results of selective excitation and saturation of a region of conformal geometric shape in two and three dimensions in a phantom as well as in vivo (healthy volunteers) are presented to demonstrate the efficacy of the method. Applications of these methods include flow- and motion-artifact suppression in body MRI, accurate conformal localization for in vivo spectroscopy in the presence of chemical shift, outer-volume suppression for resolution or speed enhancement in ultrafast imaging (Magn. Reson. Med.13, 77, 1990; J. Phys. C10, L55, 1977), and spin tagging by selective excitation/saturation of flowing spins for flow studies and angiography.     

Design of Pulse Sequences for Solvent Suppression and Uniform Excitation. Application to Observing Hemoglobin in Solution and Inside Red Blood Cells

A general method of selective excitation design for shaped pulses and pulse sequences is presented, based on analysis of the derivatives of M₊ (Δω), where M₊ is the transverse magnetization and Δω is the resonance-offset frequency. We use this method to derive pulse sequences which suppress the solvent resonance at one offset frequency and produce uniform-phase and -amplitude excitation at another offset frequency. Two families of pulse sequences are presented: small-flip-angle asymmetric pulse sequences and large-flip-angle symmetric pulse sequences. Experimental application of the pulse sequences is given for observing ¹H NMR resonances of carbonmonoxyhemoglobin and deoxyhemoglobin (deoxy-Hb) in aqueous solution and deoxy-Hb inside intact red blood cells.     

Iterative analysis of strong coupling effects in semiselective J spectroscopy

A computer program for the iterative analysis of semiselective J spectra (A. Bax, J. Magn. Reson.52, 330 (1983)) is described, and experimental and calculated f₁ cross sections through semiselective 2D J spectra for the three-spin system of fumaric acid monoethyl ester and the five-spin system of thiophene are compared.     

Binary phase shaping for selective single‐beam CARS spectroscopy and imaging of gas‐phase molecules

We report on mode‐selective single‐beam coherent anti‐Stokes Raman scattering spectroscopy of gas‐phase molecules. Binary phase shaping (BPS) is used to produce single‐mode excitation of O₂, N₂, and CO₂ vibrational modes in ambient air and gas‐phase mixtures, with high‐contrast rejection of off‐resonant Raman modes and efficient nonresonant‐background suppression. In particular, we demonstrate independent excitation of CO₂ Fermi dyads at ∼1280 and ∼1380 cm⁻¹ and apply BPS for high‐contrast imaging of CO₂ jet in ambient air. Copyright © 2010 John Wiley & Sons, Ltd.     

Single-Scan in Vivo Lactate Editing with Complete Lipid and Water Suppression by Selective Multiple-Quantum-Coherence Transfer (Sel-MQC) with Application to Tumors

A novel single-scan selective homonuclear multiple-quantum coherence-transfer technique, Sel-MQC, is presented that achieves lactate editing with complete lipid and water suppression. The method is suitable for studying tissues with high fat content and those subject to substantial motion. Frequency-selective excitation is employed to selectively prepare lactate into its multiple-quantum states; lipid and water are left in the single-quantum modes and eliminated by the multiple-quantum selection gradients. The efficiency of lipid suppression is monitored by a 2D Sel-MQC experiment which separates lipid and lactate along the multiple-quantum-evolution dimension. The spatial distribution of lactate can be imaged by the spectroscopic imaging version of Sel-MQC. Sel-MQC sequences were demonstrated both in phantoms and in vivo, using subcutaneously implanted murine EMT6 tumors.     

Coherent excitation of Rydberg atoms in an ultracold gas

We demonstrate two schemes for the coherent excitation of Rydberg atoms in an ultracold gas of rubidium atoms employing the three-level ladder system 5S_1/2-5P_3/2-n?_j. In the first approach rapid adiabatic passage with pulsed laser fields yields Rydberg excitation probabilities of 90% in the center of the laser focus. In a second experiment two-photon Rydberg excitation with continuous-wave fields is applied which results in Rabi oscillations between the ground and Rydberg state. The experiments represent a prerequisite for the control of interactions in ultracold Rydberg gases and the application of ultracold Rydberg gases for quantum information processing.     

A heteronuclear intermolecular single-quantum coherence scheme for high-resolution 2D J-resolved 1H NMR spectra in inhomogeneous magnetic fields

Based on heteronuclear intermolecular single-quantum coherences between proton (¹H) and quadrupolar nuclei (i.e. deuterium ²H), a three-dimensional nuclear magnetic resonance (NMR) pulse sequence is proposed for recovering high-resolution two-dimensional J-resolved NMR spectra from samples mixed with a deuterated solvent in the presence of large magnetic field inhomogeneities. Benefitting from excitation of spins via two different radio frequency (RF) transmit channels, this sequence is suitable for applications in randomly large inhomogeneous fields and the solvent suppression generally required in homonuclear intermolecular multiple-quantum coherence approaches is no longer necessary. Systematic theoretical analyses are given based on the distant dipolar field treatment. Experiment on a sample of corn oil in deuterated acetone and ethyl 3-bromopropionate and acetone dissolved in DMSO-d₆ in a deshimmed field with severe inhomogeneous broadening is performed to show the feasibility and applicability of this sequence.     

Complete NMR fingerprints of proteins in H2O solution without solvent presaturation. An application of two-dimensional longitudinal two-spin-order spectroscopy

A method for obtaining complete two-dimensional ¹H NMR amide proton-C_α proton J-connectivity maps for proteins (the “fingerprint”) is describe. The method relies on the conversion of antiphase single-quantum coherence into longitudinal two-spin order which is made observable by utilizing a selective solvent-suppression readout pulse. Since irradiation of the solvent resonance is unnecessary in this experiment, J connectivities are observable even for protons resonating exactly at the solvent frequency. Thus, a complete protein fingerprint is obtained at a single set of experimental conditions. The method is demonstrated for a 75-amino-acid protein.     

Hadamard excitation sculpting.

An approach to Hadamard phase encoding and editing in an excitation sculpting experiment is presented. When band- and/or frequency-selective experiments are performed at more than one site using excitation sculpting, use of Hadamard excitation sculpting (HEX sculpting) will reduce the total measuring time to achieve a target S/N. The application of HEX sculpting is demonstrated using selective 1D and NOESY1D experiments.     

Solvent suppression using phase-modulated binomial-like sequences and applications to diffusion measurements

Two phase-modulated binomial-like pi pulses have been developed by simultaneously optimizing pulse durations and phases. In combination with excitation sculpting, both of the new binomial-like sequences outperform the well-known 3-9-19 sequence in selectivity and inversion width. The new sequences provide similar selectivity and inversion width to the W5 sequence but with significantly shorter sequence durations. When used in PGSTE-WATERGATE, they afford highly selective solvent suppression in diffusion experiments.     

Convection-compensating PGSE experiment incorporating excitation-sculpting water suppression (CONVEX)

We present a new diffusion experiment which provides simultaneous suppression of an on-resonance solvent peak and compensation for convection. The experiment, which we name CONVEX, exploits similarities between two functionally different pulse sequences to enable the same sequence to be used simultaneously for two different purposes. The CONVEX pulse sequence combines a double-echo PGSE with double excitation-sculpting water suppression, using unequal gradient pulse-pair amplitudes (g1 and g2) and unequal diffusion intervals (Delta1 and Delta2). Convection compensation is achieved by setting g1:g2 = Delta2:Delta1. The new experiment provides the spectral quality, flat baseline, and water-suppression power characteristic of excitation-sculpting experiments, combined with excellent compensation for convection. The resulting Stejskal-Tanner plots are linear over a greater range of signal attenuation than in the absence of water suppression. Possible applications include protein NMR; NMR of cellular or colloidal systems; and the monitoring of technological processes.