To analyze the intracellular chemical conditions of living cells, the sample is frozen until required and then treated with a low melting point solvent for spectrometry using NMR

摘要

To analyze the intracellular chemical condition of living cells, using nuclear magnetic resonance (NMR), the sample is immersed directly and immediately into liquid nitrogen to freeze the sample rapidly. The sample is stored at a very low temperature. At the time required for analysis, the sample is treated with a solvent with a very low melting point. To analyze the intracellular chemical condition of living cells, a mixture of water and methanol can be used instead of liquid nitrogen, plugged at a very low temperature at a pH of 7.5 and with a 50% methanol content. The solvent is a mixture of deutered water and deutered methanol, at a pH of 7.5 and with a 50% methanol content. The low temperature used is <= 0 deg C or -10 deg C to -80 deg C or <= -80 deg C. During the NMR analysis, the temperature is <= 0 deg C or -5 deg C to -20 deg C. The analysis is effected by NMR with high resolution magic angle spectrometry (HRMAS) and preferably in two dimensions. The sample is taken in a cellular culture environment or tissue, containing molecules enriched with stable isotopes where the living cells are cultivated or incubated. The molecules contain a source of carbon 13 and/or nitrogen 15. The living cells are prokaryote or eukaryote animal or vegetable cells. Independent claims are included for the following: (1) an application of the living cell analysis, to determine deviations from a normal physiology; and (2) an apparatus with a superconductor magnet, an electronic control unit, a NMR probe and a freezing unit.