CEA Saclay, DSM, IRAMIS, UMR CEA/CNRS 3299 - NIMBE, Laboratoire Structure et Dynamique par Résonance Magnétique Gif-sur-Yvette, France.
Department of Chemistry, University of York Heslington, York, UK.
Front Chem. 2014 Jun 12;2:38. doi: 10.3389/fchem.2014.00038. eCollection 2014.
The low sensitivity and thus need for large sample volume is one of the major drawbacks of Nuclear Magnetic Resonance (NMR) spectroscopy. This is especially problematic for performing rich metabolic profiling of scarce samples such as whole cells or living organisms. This study evaluates a (1)H HR-MAS approach for metabolic profiling of small volumes (250 nl) of whole cells. We have applied an emerging micro-NMR technology, high-resolution magic-angle coil spinning (HR-MACS), to study whole Saccharomyces cervisiae cells. We find that high-resolution high-sensitivity spectra can be obtained with only 19 million cells and, as a demonstration of the metabolic profiling potential, we perform two independent metabolomics studies identifying the significant metabolites associated with osmotic stress and aging.
低灵敏度和因此需要大样本量是核磁共振(NMR)光谱学的主要缺点之一。对于对稀缺样本(如整个细胞或生物体)进行丰富的代谢分析来说,这是一个特别成问题的问题。本研究评估了(1)H HR-MAS 方法在小体积(250 nl)整个细胞代谢分析中的应用。我们已经应用了新兴的微 NMR 技术,高分辨率魔角旋转(HR-MACS),来研究整个酿酒酵母细胞。我们发现,仅用 1900 万个细胞就可以获得高分辨率、高灵敏度的光谱,并且作为代谢分析潜力的证明,我们进行了两项独立的代谢组学研究,确定了与渗透压应激和衰老相关的重要代谢物。