Cryogenic sample loading into a magic angle spinning nuclear magnetic resonance spectrometer that preserves cellular viability

Rupam Ghosh, Jaka Kragelj, Yiling Xiao, Kendra K. Frederick

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Dynamic nuclear polarization (DNP) can dramatically increase the sensitivity of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. These sensitivity gains increase as temperatures decrease and are large enough to enable the study of molecules at very low concentrations at the operating temperatures (~100 K) of most commercial DNP-equipped NMR spectrometers. This leads to the possibility of in-cell structural biology on cryopreserved cells for macromolecules at their endogenous levels in their native environments. However, the freezing rates required for cellular cryopreservation are exceeded during typical sample handling for DNP MAS NMR and this results in loss of cellular integrity and viability. This article describes a detailed protocol for the preparation and cryogenic transfer of a frozen sample of mammalian cells into a MAS NMR spectrometer.

Original languageEnglish (US)
Article numbere61733
Pages (from-to)1-12
Number of pages12
JournalJournal of Visualized Experiments
Volume2020
Issue number163
DOIs
StatePublished - 2020

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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