benchtop spinsolve ultra 60 mhz nmr spectrometer  (Magritek Ltd)

Journal: Analytical Chemistry

Article Title: Robust Rapid Cellular Metabolite Sensing Using Benchtop NMR and SABRE-Hyperpolarized [1- 13 C]Pyruvate

doi: 10.1021/acs.analchem.5c05076

Figure Lengend Snippet: Graphical workflow for benchtop assessment of cellular metabolic flux with SABRE hyperpolarization. [A] Graphical depiction of the cell preparation process, where different cell subtypes could be used with the depicted benchtop workflow. [B] Bubbling setup used for SABRE hyperpolarization, where parahydrogen gas is regulated through an NMR tube in a shielded magnetic environment with an applied external field (typically 0.4 μT). [C] SABRE hyperpolarization chemistry, where parahydrogen and pyruvate exchange reversibly on an iridium metal catalyst (here, Ir-IMes), generating HP [1- 13 C]­pyruvate from transfer of the parahydrogen spin order to the pyruvate 13 C nuclear spin. [D] Benchtop SABRE polarizer and NMR spectrometer, where injection of cells and HP pyruvate generates a time series of metabolic spectra. [E] Products of HP [1- 13 C]­pyruvate, showing the conversion of [1- 13 C]­pyruvate to [1- 13 C]­alanine via alanine aminotransferase (AAT) or shuttling into the mitochondria and TCA cycle for subsequent oxidative decarboxylation via aerobic metabolism (transport protein mitochondrial pyruvate carrier, MPC, and pyruvate dehydrogenase complex, PDH).

Article Snippet: Three minutes prior to an experiment where yeast metabolic flux was measured, 0.3 mL aliquots of the preincubated yeast suspension were transferred into 35 °C preheated high-throughput 5 mm NMR tubes (Wilmad Labglass) and placed into a 1.4 T benchtop NMR spectrometer (SpinSolve Carbon, Magritek).

Techniques: Injection

Journal: Analytical Chemistry

Article Title: Robust Rapid Cellular Metabolite Sensing Using Benchtop NMR and SABRE-Hyperpolarized [1- 13 C]Pyruvate

doi: 10.1021/acs.analchem.5c05076

Figure Lengend Snippet: [A] Spectra from three separate SABRE HP [1- 13 C]­pyruvate samples, showing polarization both under hydrogen pressure, in situ in the 5 mm NMR tube reactor (left) and after processing for each respective sample (right). The difference in spectral line width between the in situ (left) and processed (right) spectra is potentially due to the dynamic exchange of the SABRE process and potential susceptibility artifacts from the bubbling catheter in the in situ sample, while the purified sample experiences no SABRE exchange. [B] Spectrum from a pure [1- 13 C]­acetic acid reference sample (17.18 M) acquired in a single shot. All spectra in [A] and [B] were acquired with a 7° flip angle on a 1.4 T SpinSolve Carbon benchtop NMR (Magritek). [C] Relaxation measurements in situ (in the hyperpolarization solution, under p -H 2 pressure) at seven different magnetic fields. [D] Purification process used to transfer SABRE HP [1- 13 C]­pyruvate from an 80% acetone in D 2 O mixture into an aqueous solution, where the magnet loop represents the 0.2 T Halbach array.

Article Snippet: Three minutes prior to an experiment where yeast metabolic flux was measured, 0.3 mL aliquots of the preincubated yeast suspension were transferred into 35 °C preheated high-throughput 5 mm NMR tubes (Wilmad Labglass) and placed into a 1.4 T benchtop NMR spectrometer (SpinSolve Carbon, Magritek).

Techniques: In Situ, Purification