Weybright P, Millis K, Campbell N, Cory D G, Singer S
Department of Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Magn Reson Med. 1998 Mar;39(3):337-45. doi: 10.1002/mrm.1910390302.
The application of new gradient, high-resolution, magic angle spinning (MAS) 1H nuclear magnetic resonance (NMR) spectroscopy to the study of intact undifferentiated and differentiated NIH 3T3 F442A cells demonstrated improved spectral resolution and sensitivity compared with static studies. MAS of cells permits the detection and quantitation of many cellular metabolites that are not clearly resolved in nonspinning measurements and provides an improved visibility of phospholipids. Gradient, MAS enables the use of diffusion weighting for compartment assignment and the determination of mobility for many metabolites which are incompletely resolved using static techniques. The smaller, undifferentiated preadipocytes show no microscopic evidence of cell lysis after 2 h of MAS at 3.5 kHz and 82% of these cells remain viable by trypan blue exclusion. In contrast, 15-19% of the larger, lipid-laden differentiated adipocytes were found to suffer some degree of cell lysis with MAS. This new method is an attractive alternative to either nonspinning perfusion or extraction techniques for NMR studies of cells.
将新型梯度、高分辨率、魔角旋转（MAS）1H核磁共振（NMR）光谱应用于完整的未分化和分化的NIH 3T3 F442A细胞研究，结果表明与静态研究相比，其光谱分辨率和灵敏度有所提高。对细胞进行MAS能够检测和定量许多在非旋转测量中无法清晰分辨的细胞代谢物，并能更好地观察磷脂。梯度MAS能够利用扩散加权进行区室分配，并确定许多使用静态技术无法完全分辨的代谢物的流动性。较小的未分化前脂肪细胞在3.5 kHz下进行2小时的MAS后，未显示出细胞裂解的微观证据，并且通过台盼蓝排斥法检测发现其中82%的细胞仍具有活力。相比之下，发现较大且富含脂质的分化脂肪细胞中有15 - 19%在进行MAS时会出现一定程度的细胞裂解。这种新方法是细胞NMR研究中替代非旋转灌注或提取技术的一种有吸引力的选择。
