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Hyperpolarization-Enhanced NMR Spectroscopy with Femtomole Sensitivity Using Quantum Defects in Diamond

MetadataDetails
Publication Date2020-06-09
JournalPhysical Review X
AuthorsDominik Bucher, David R. Glenn, Hongkun Park, Mikhail D. Lukin, Ronald L. Walsworth
InstitutionsCenter for Astrophysics Harvard & Smithsonian, Technical University of Munich
Citations83
AnalysisFull AI Review Included
  • Core Achievement: Successful integration of Overhauser Dynamic Nuclear Polarization (DNP) with picoliter-scale Nitrogen-Vacancy (NV) ensemble NMR spectroscopy (DNP-NV-NMR).
  • Sensitivity Breakthrough: Achieved femtomole (50 fmol) molecule-number sensitivity, representing a greater than two orders of magnitude improvement over previous NV-NMR methods.
  • Performance Metrics: Demonstrated a proton number sensitivity of ~10 pmol/(Hz)1/2, enabling high-resolution spectroscopy on dilute solutions.
  • Enhancement Factor: Observed a ~230x signal enhancement in water samples due to DNP using TEMPOL molecular radicals at a low bias field (84.7 mT).
  • High Resolution: Maintained high spectral resolution (~8-10 Hz linewidths) sufficient to observe chemical shifts and J-couplings in various small organic molecules (e.g., xylene, thymine).
  • Technical Implementation: The system uses a 12C-enriched CVD diamond chip with a 13 ”m 14N-doped surface layer, defining a small sensing volume (~10 pL). The existing GHz-frequency antenna is leveraged for both NV spin manipulation and DNP driving.
  • Impact: The technique advances mass-limited NMR spectroscopy, opening pathways for applications in single-cell analysis, metabolomics, and high-throughput drug screening.
ParameterValueUnitContext
Molecule Number Sensitivity (LOD)50femtomoleLimit of detection (SNR=3) for t-BuOD
Proton Number Sensitivity10pmol/(Hz)1/2Hyperpolarized water sample (SNR=3)
DNP Signal Enhancement~230xCompared to non-DNP control
Effective NMR Sensing Volume~10pLDefined by laser spot size on diamond
Bias Magnetic Field (B0)84.7mTOperating field for NV-NMR
NV Resonance Frequency~500MHzElectron spin transition frequency
TEMPOL ESR Frequency~2.37GHzOverhauser drive frequency
Proton NMR Frequency~3.606MHzFNP detection frequency
NV Ensemble Density~3x1017cm-3In the 13 ”m surface layer
Surface Layer Thickness13”m14N-enriched layer on diamond
Diamond T2* (Ramsey)~750nsNV ensemble coherence time
Diamond T2 (Hahn Echo)~6.5”sNV ensemble coherence time
Proton Spin Lifetime (T1)~150msWith 20 mM TEMPOL radical
Laser Wavelength (λ)532nmGreen laser for NV readout
Laser Spot Diameter~20”mDefining active sensor area
Magnetometer Noise Floor~20pT/Hz-1/2NV ensemble sensor performance
Spectral Linewidth (Δf)8-10HzObserved in molecular spectra
Maximum DNP Rabi Frequency~10MHzOptimized drive power for DNP

The DNP-enhanced NV-NMR system relies on combining specialized diamond fabrication, precise microwave control, and a synchronized readout sequence.

  1. Diamond Sensor Preparation:

    • Substrate: 12C enriched (99.999%) Chemical Vapor Deposition (CVD) diamond chip (2mm x 2mm x 0.5 mm).
    • NV Layer Growth: A 13 ”m thick surface layer was grown with high 14N density (~4.8 x 1018 cm-3).
    • NV Creation: Electron irradiation (flux 1.3 x 1014 cm-2 s-1) followed by annealing at 800 °C yielded a dense NV ensemble (~3x1017 cm-3).
    • Optical Geometry: Diamond edges were polished at 45° to enable laser excitation via total internal reflection (TIR), minimizing light exposure to the liquid sample.
  2. Magnetic and Microwave Setup:

    • Bias Field (B0): A feedback-stabilized electromagnet provided B0 = 84.7 mT, aligned parallel to the diamond’s [111] axis.
    • Antenna: A 1 mm diameter wire loop antenna, mounted immediately above the diamond, was used to drive both the NV spins (~500 MHz) and the DNP agent spins (~2.37 GHz).
    • NMR Drive: A separate resonant coil (Q ~140) was used to apply the π/2 pulse on the protons at ~3.606 MHz, achieving a proton Rabi frequency of ~4 kHz.
  3. Hyperpolarization (Overhauser DNP):

    • Agent: TEMPOL (4-Hydroxy-TEMPO) molecular radicals were dissolved in the liquid sample (e.g., 20 mM in water).
    • Process: Continuous microwave driving saturates the TEMPOL electronic spin transition (at 2.37 GHz), transferring thermal electron spin polarization to the nuclear spins of the sample molecules (protons).
  4. Detection Sequence (DNP-CASR):

    • The full experiment alternates between the Overhauser DNP sequence (~2 x T1, typically 300 ms) and the FNP detection sequence (~4 x T2*, typically 200 ms).
    • FNP Generation: A π/2 pulse is applied to the hyperpolarized proton spins.
    • CASR Readout: The Free Nuclear Precession (FNP) signal is detected by the NV ensemble using the Coherently Averaged Synchronized Readout (CASR) magnetometry pulse sequence, which is based on XY8-6 dynamic decoupling subsequences synchronized to an external clock.

The combination of femtomole sensitivity and picoliter volume selectivity makes this technology highly relevant for mass-limited analytical fields:

  • Drug Discovery and Screening:
    • High-throughput screening (HTS) of binding affinity for mass-limited chemical reactions (nanomole scale synthesis).
    • Providing superior isomeric distinguishability compared to traditional mass spectrometry pipelines.
  • Metabolomics and Cell Biology:
    • Quantitative metabolic studies and real-time analysis of metabolic flux at the single-cell level, leveraging the picoliter volume selectivity.
    • Analysis of small, precious biological samples where sample volume is severely restricted.
  • Catalysis and Chemical Synthesis:
    • High-throughput analysis of mass-limited chemical reactions, allowing rapid characterization of intermediates and products using minimal reagent quantities.
  • Advanced Magnetic Resonance Imaging (MRI):
    • Enabling NV-detected MRI techniques at the micrometer scale, potentially allowing studies of water diffusion and transport dynamics within cells and tissue.
  • Quantum Sensing Technology:
    • Development of next-generation, ultra-sensitive solid-state NMR sensors capable of operating at small bias fields (85 mT) while achieving high sensitivity typically associated with high-field inductive detectors.
View Original Abstract

Nuclear magnetic resonance (NMR) spectroscopy is a widely used tool for chemical analysis and molecular structure identification. Because it typically relies on the weak magnetic fields produced by a small thermal nuclear spin polarization, NMR suffers from poor molecule-number sensitivity compared to other analytical techniques. Recently, a new class of NMR sensors based on optically-probed nitrogen-vacancy (NV) quantum defects in diamond have allowed molecular spectroscopy from sample volumes several orders of magnitude smaller than the most sensitive inductive detectors. To date, however, NV-NMR spectrometers have only been able to observe signals from pure, highly concentrated samples. To overcome this limitation, we introduce a technique that combines picoliter-scale NV-NMR with fully integrated Overhauser dynamic nuclear polarization (DNP) to perform high-resolution spectroscopy on a variety of small molecules in dilute solution, with femtomole sensitivity. Our technique advances mass-limited NMR spectroscopy for drug and natural product discovery, catalysis research, and single cell studies.