Slow-spinning low-sideband HR-MAS NMR spectroscopy: delicate analysis of biological samples

Renault, Marie; Shintu, Laetitia; Piotto, Martial; Caldarelli, Stéfano

doi:10.1038/srep03349

Cited by 28 publications

(30 citation statements)

References 34 publications

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“…The g-forces associated with the advocated sample packing approach can exceed 100,000 × g (Table 1), which may indeed lead to irrevocable changes in certain samples (Han et al 2010; Renault et al 2013; Cai et al 2014). Whilst this is not an unreasonable concern, it is important to realize that this may be a moot point when samples are used for MAS ssNMR.…”

Section: Discussionmentioning

confidence: 99%

On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR

et al. 2017

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A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation.

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Section: Discussionmentioning

confidence: 99%

On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR

et al. 2017

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“…Recently, Caldarelli and coworkers 59,60 demonstrated that high-quality 1 H spectra can be acquired at spinning frequencies of a few hundred hertz without the need for complex sideband suppression pulse sequences. The idea is to reduce the sideband intensities by minimizing the susceptibility gradient within the sample.…”

Section: Slow Mas Experimentsmentioning

confidence: 99%

High-resolution Magic-angle Spinning (HR-MAS) NMR Spectroscopy

Wong

Lucas‐Torres

2018

NMR-based Metabolomics

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Since the beginning of high-resolution magic-angle spinning (HR-MAS) NMR spectroscopy in 1990s, we have witnessed tremendous instrumentation and methodological advancements in the HR-MAS NMR technique for semisolids. With HR-MAS, it is now possible to acquire reliable high-quality spectra in a routine and high-throughput fashion, and it has become a well-integrated metabolic screening tool for ex vivo biospecimens such as tissue biopsies, cells and organisms for NMR-based metabolomics research. This chapter provides the basic principles of HR-MAS and describes a few recent noteworthy developments that could strengthen the role of HR-MAS as a frontline NMR technique for metabolomics.

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“…Figure 1 shows a six-step protocol that builds upon observations made with SSBreduced slow-HRMAS experiments and introduces several new, essential steps. 20,25,26 Figures 3 and 4 show the 1D 1 H HR-MAS NMR spectra, obtained at 4000 Hz MAS, of a heifer liver sample contained either in a 12-μL rotor (Figures 3a and b) or in a disposable insert (Figures 4a and b). The spectrum consists of an envelope of broad signals that could arise either from macromolecules and from components with restricted molecular motions superimposed on well-resolved peaks from small molecules characterized by fast and isotropic mobility.…”

Section: Samplesmentioning

confidence: 99%

“…20 These studies are motivated by the expected source of SSB in HR-MAS for the mobile metabolites. In biological samples, which contain many intracellular and intercellular structures, metabolites are distributed heterogeneously, and often close to a boundary of a material with a different susceptibility than the compartment containing the compound under observation.…”

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confidence: 99%

Complete Protocol for Slow-Spinning High-Resolution Magic-Angle Spinning NMR Analysis of Fragile Tissues

et al. 2014

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High-resolution magic-angle spinning (HR-MAS) nuclear magnetic resonance (NMR) is an essential tool to characterize a variety of semisolid systems, including biological tissues, with virtually no sample preparation. The "non-destructive" nature of NMR is typically compromised, however, by the extreme centrifugal forces experienced under conventional HR-MAS frequencies of several kilohertz. These features limit the usefulness of current HR-MAS approaches for fragile samples. Here, we introduce a full protocol for acquiring high-quality HR-MAS NMR spectra of biological tissues at low spinning rates (down to a few hundred hertz). The protocol first consists of a carefully designed sample preparation, which yields spectra without significant spinning sidebands at low spinning frequency for several types of sample holders, including the standard disposable inserts classically used in HR-MAS NMR-based metabolomics. Suppression of broad spectral features is then achieved using a modified version of the recently introduced PROJECT experiment with added water suppression and rotor synchronization, which deposits limited power in the sample and which can be suitably rotor-synchronized at low spinning rates. The performance of the slow HR-MAS NMR procedure is demonstrated on conventional (liver tissue) and very delicate (fish eggs) samples, for which the slow-spinning conditions are shown to preserve the structural integrity and to minimize intercompartmental leaks of metabolites. Taken together, these results expand the applicability and reliability of HR-MAS NMR spectroscopy. These results have been obtained at 400 and 600 MHz and suggest that high-quality slow HR-MAS spectra can be expected at higher magnetic fields using the described protocol.

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Slow-spinning low-sideband HR-MAS NMR spectroscopy: delicate analysis of biological samples

Cited by 28 publications

References 34 publications

On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR

On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR

High-resolution Magic-angle Spinning (HR-MAS) NMR Spectroscopy

Complete Protocol for Slow-Spinning High-Resolution Magic-Angle Spinning NMR Analysis of Fragile Tissues

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