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Avance 3 nmr console

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The Avance III NMR console is a modular and highly flexible platform for nuclear magnetic resonance (NMR) spectroscopy. It provides the core functionality required to operate an NMR spectrometer and acquire high-quality data.

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Lab products found in correlation

Er 4119hs lc sensitivity cavity, by Bruker (2 mentions) Emxplus spectrometer, by Bruker (2 mentions) Er 4119hs lc, by Bruker (1 mentions) Emx cw epr spectrometer, by Bruker (1 mentions) 10 mm cryoprobe, by Bruker (1 mentions) Topspin, by Bruker (1 mentions)

6 protocols using avance 3 nmr console

1

Hyperpolarized Pyruvate NMR Spectroscopy

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Spectra in HP pyruvate experiments were collected in a custom-built 20 mm broadband probe (QOneTec, Switzerland) installed in a 14 T magnet equipped with an Avance III NMR console (Bruker Biospin, United States). Shimming was carried out using the 23Na signal prior to the HP experiment. Linewidths of ~18 Hz in 23Na spectrum were achieved. 13C spectra (spectral width of 300 ppm; 32,768 data points) were recorded using 30° radiofrequency pulses with 1H decoupling (WALTZ65) during acquisition and a repetition time of 3 s. Spectra were processed with 20 Hz exponential line broadening and baseline corrected using a polynomial function. Peak areas of metabolites were obtained by fitting mixed Lorentzian-Gaussian line shapes. NMR spectra were processed using MestReNova (v14.2.1).
Ragavan M., McLeod M.A., Rushin A, & Merritt M.E. (2022). Detecting de novo Hepatic Ketogenesis Using Hyperpolarized [2-13C] Pyruvate. Frontiers in Physiology, 13, 832403.
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2

ODNP Analysis of Spin-Labeled Samples

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Samples of approximately 3.5 µL volume were placed in a 0.6 mm i.d. and 0.84 o.d. quartz capillary and analyzed by ODNP, as described previously21 (link). A “pass through” NMR probe design built to fit inside a 3mm i.d. 6 mm o.d. quartz tube was used. The quartz tube was inserted into a high sensitivity microwave cavity (ER 4119HS-LC, Bruker Biospin) along with the NMR probe and sample. ODNP experiments were performed using a Bruker EMX CW EPR spectrometer and a Bruker Avance III NMR console. The samples were sealed in a capillary with a protective layer of Critoseal on the top and hot beeswax on the bottom. All ODNP measurements were performed at room temperature. The sample was irradiated with up to 6 W of microwaves at the EPR resonant frequency of the spin labels at ~10 GHz using a home-built microwave amplifier22 (link). The magnetic field was set on resonance at the central electron hyperfine transition, here at 9.8 GHz. The spin label concentration of each sample was determined from the double integral of its cw EPR spectrum measured at 1 mW irradiation power, 0.4 G modulation width. The concentration-dependent ODNP relaxation rates, kσ, kρ, and kLow, are normalized to the sample concentration derived from spin counting per integration of the cw EPR spectrum.
Barnes R., Sun S., Fichou Y., Dahlquist F.W., Heyden M, & Han S. (2017). Spatially heterogeneous surface water diffusivity around structured protein surfaces at equilibrium. Journal of the American Chemical Society, 139(49), 17890-17901.
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3

Overhauser Dynamic Nuclear Polarization Technique

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Overhauser dynamic nuclear polarization was performed on samples (3.5 μL) that contained spin-labeled ɛPL. The samples were loaded into quartz capillaries of 0.6 mm ID × 0.84 mm OD (Vitrocom, New Jersey, USA), and both ends of the tubes were sealed with Critoseal. ODNP experiments were performed using a Bruker EMXPlus spectrometer and a Bruker Avance III NMR console (Bruker, Massachusetts, USA). The capillary tube was mounted on a home-built NMR probe with a U-shaped NMR coil, and was set in a Bruker ER 4119HS-LC sensitivity cavity. Samples were irradiated at 9.8 GHz with the center field set at 3484 G and sweep width of 120 G. Dry air was streamed through the NMR probe during all measurements. Theory of ODNP and details in the experiment are previously reported in other studies65 (link)–67 (link).
Park S., Barnes R., Lin Y., Jeon B.J., Najafi S., Delaney K.T., Fredrickson G.H., Shea J.E., Hwang D.S, & Han S. (2020). Dehydration entropy drives liquid-liquid phase separation by molecular crowding. Communications Chemistry, 3, 83.
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4

Hyperpolarized 13C NMR Spectroscopy of DHA

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Spectra in hyperpolarized DHA experiments were collected in a custom-built 20 mm broadband probe (Qonetec, Dietlikon, Switzerland) installed in a 14 T magnet equipped with an Avance III NMR console (Bruker Biospin, Billerica, MA, USA). Prior to injection of HP DHA, shimming was carried out using the 23Na signal. Typically, linewidths of around 18 Hz in 23Na spectrum were achieved (translating to similar linewidths in 13C spectra). 13C spectra (spectral width of 424 ppm; 12,280 data points) were recorded using a 45° {x, −x} binomial excitation scheme centered on DHA resonance (nominally, 212 ppm) with 1H decoupling (WALTZ65; B1 = 4.5 KHz) during acquisition and a repetition time of 3 s. Spectra were processed with 15–20 Hz exponential line broadening and baseline corrected using a polynomial function. Peak areas of metabolites were obtained by fitting mixed Lorentzian-Gaussian line shapes to 13C sum spectra (i.e., sum of individual 13C spectra acquired post DHA administration). We do not include 2H NMR spectra of the glucose in our analysis, as we achieved unexpectedly high enrichments of both 13C and 2H. Excessive 13C enrichment rendered the 2H spectra unusable due to overlap between the peaks and adjacent 13C j-coupled satellites.
Ragavan M., McLeod M.A., Giacalone A.G, & Merritt M.E. (2021). Hyperpolarized Dihydroxyacetone Is a Sensitive Probe of Hepatic Gluconeogenic State. Metabolites, 11(7), 441.
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5

Hyperpolarized 13C NMR Spectroscopy of Perfused Pancreata

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Spectra of the perfusing pancreata were collected in 10 mm cryoprobe (Bruker Biospin, Billerica, MA, USA) installed in a 14-T magnet with an Avance III NMR console (Bruker Biospin, USA). Shimming was carried out using the 23Na signal with final linewidths of ~17 Hz. 13C spectra (spectral width of 300 ppm; 32,768 data points) were recorded using 15° radiofrequency pulses and 1H decoupling (WALTZ65) during acquisition with a 3-s repetition time. 10 Hz exponential line broadening and baseline correction using a third-order polynomial function were applied to all spectra. The kinetics of individual metabolites were measured using automatic integration in Bruker TopSpin (v4.1.3) across individual spectra followed by total carbon normalization. Summed spectra were processed in MestReNova (v14.2.1), and peak areas of metabolites were obtained by fitting mixed Lorentzian–Gaussian line shapes. The signals were normalized to the total carbon intensity of a sum spectrum of the 60 data points in time where lactate was present. Resonances were assigned using known chemical shifts and previously reported work with hyperpolarized substrates and ex vivo perfusions.24 ,25 (link) Chemical shift assignments were confirmed with [1H, 13C] HSQC and [1H, 13C] HMBC experiments.
Rushin A., McLeod M.A., Ragavan M, & Merritt M.E. (2023). Observing exocrine pancreas metabolism using a novel pancreas perfusion technique in combination with hyperpolarized [1-13C]pyruvate. Magnetic resonance in chemistry : MRC, 61(12), 748-758.
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6

ODNP Analysis of Spin-Labeled Tau187

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We used the aforementioned four spin labeled tau187 variants (singly labeled on site 303, 313, 322 and 404, respectively), and followed previously reported methods for ODNP experiments [30 (link)]. 100 μM spin labeled tau187 was mixed with designated concentration of NaCl. Immediately after sample preparation, 3.5 μL of the samples were loaded into quartz capillaries of 0.6 mm ID × 0.84 mm OD (Vitrocom, New Jersey, USA), and two ends of the tubes were sealed with Critoseal and beeswax respectively. ODNP experiments were performed using a Bruker EMXPlus spectrometer and a Bruker Avance III NMR console (Bruker, Massachusetts, USA). The capillary tube was mounted on a home-built NMR probe with a U-shaped NMR coil, and was set in a Bruker ER 4119HS-LC sensitivity cavity. Samples were irradiated at 9.8 GHz with the center field set at 3484 G and sweep width of 120 G. Dry air was kept at temperature of 18 °C and purged through the NMR probe during all measurements. ODNP data was analyzed using customized python script WorkupODNP and ODNPLab, available on https://www.github.com. Theory of ODNP and details in the experiment are previously reported in other studies [63 (link)], [64 (link)].
Lin Y., Fichou Y., Longhini A.P., Llanes L.C., Yin P., Bazan G.C., Kosik K.S, & Han S. (2020). Liquid-liquid phase separation of tau driven by hydrophobic interaction facilitates fibrillization of tau. Journal of molecular biology, 433(2), 166731.
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