CEA, IRAMIS, SIS2M, Laboratoire Structure et Dynamique par Résonance Magnétique, Gif-sur-Yvette, France.
NMR Biomed. 2011 Dec;24(10):1264-9. doi: 10.1002/nbm.1686. Epub 2011 Mar 15.
We show that the differentiation between internal and external compartments of various biological cells in suspension can be made via simple NMR spectra of hyperpolarized (129) Xe. The spectral separation between the signals of (129) Xe in these two compartments is already known for red blood cells, because of the strong interaction of the noble gas with hemoglobin. The observation of two separate peaks in the 200-ppm region can be seen with both eukaryotic and prokaryotic cells, some of which are not known to contain paramagnetic proteins in large quantities. Using different experiments in which the cells are lysed, swell or are blocked in G2 phase, we demonstrate that the low-field-shifted peak observed corresponds to xenon in the aqueous pool inside the cells and not in the membranes. The presence of this additional peak is a clear indication of cell integrity, and its integration allows the quantification of the total cell volume. The relaxation time of intracellular xenon is sufficiently long to open up promising perspectives for cell characterization. The exchange time between the inner and outer cell compartments (on the order of 30 ms) renders possible the targeting of intracellular receptors, whereas the observation of chemical shift variations represents a method of revealing the presence of toxic species in the cells.
我们表明,通过对超极化(129)Xe 的简单 NMR 谱,可以区分悬浮的各种生物细胞的内部和外部隔室。由于稀有气体与血红蛋白的强烈相互作用,已经知道(129)Xe 在这两个隔室中的信号之间存在光谱分离。可以在真核细胞和原核细胞中观察到 200 ppm 区域中两个单独的峰,其中一些细胞不含有大量的顺磁蛋白。通过对细胞裂解、肿胀或在 G2 期受阻的不同实验的观察,我们证明观察到的低场移动峰对应于细胞内水溶液中的氙气,而不是在细胞膜中。存在这个额外的峰是细胞完整性的明确指示,其积分允许定量细胞的总体积。细胞内氙的弛豫时间足够长,为细胞特征化开辟了有希望的前景。内外细胞隔室之间的交换时间(约 30 ms)使得靶向细胞内受体成为可能,而观察化学位移变化则代表了揭示细胞中有毒物质存在的方法。