Bastawrous Monica, Ghosh Biswas Rajshree, Soong Ronald, Jouda Mazin, MacKinnon Neil, Mager Dario, Korvink Jan G, Simpson Andre J
Environmental NMR Center, University of Toronto, 1265 Military Trail, Toronto, Ontario M1C 1A4, Canada.
Institute of Microstructure Technology, Karlsruhe Institute of Technology, 76344 Eggenstein-Leopoldshafen, Germany.
Anal Chem. 2023 Jan 17;95(2):1327-1334. doi: 10.1021/acs.analchem.2c04203. Epub 2022 Dec 28.
Nuclear magnetic resonance (NMR) spectroscopy is commonly employed in a wide range of metabolomic research. Unfortunately, due to its relatively low sensitivity, smaller samples become challenging to study by NMR. Cryoprobes can be used to increase sensitivity by cooling the coil and preamplifier, offering sensitivity improvements of ∼3 to 4. Alternatively, microcoils can be used to increase mass sensitivity by improving sample filling and proximity, along with decreased electrical resistance. Unfortunately, combining the two approaches is not just technically challenging, but as the coil decreases, so does its thermal fingerprint, reducing the advantage of cryogenic cooling. Here, an alternative solution is proposed in the form of a Lenz lens inside a cryoprobe. Rather than replacing the detection coil, Lenz lenses allow the B field from a larger coil to be refocused onto a much smaller sample area. In turn, the stronger B field at the sample provides strong coupling to the cryocoil, improving the signal. By combining a 530 I.D. Lenz lens with a cryoprobe, sensitivity was further improved by 2.8 and 3.5 for H and C, respectively, over the cryoprobe alone for small samples. Additionally, the broadband nature of the Lenz lenses allowed multiple nuclei to be studied and heteronuclear two-dimensional (2D) NMR approaches to be employed. The sensitivity improvements and 2D capabilities are demonstrated on 430 nL of hemolymph and eight eggs (∼350 μm O.D.) from the model organism. In summary, combining Lenz lenses with cryoprobes offers a relatively simple approach to boost sensitivity for tiny samples while retaining cryoprobe advantages.
核磁共振(NMR)光谱法在广泛的代谢组学研究中普遍应用。遗憾的是,由于其灵敏度相对较低,对于较小的样本,用NMR进行研究颇具挑战。低温探头可通过冷却线圈和前置放大器来提高灵敏度,灵敏度可提高约3至4倍。另外,微线圈可通过改善样品填充和接近度以及降低电阻来提高质量灵敏度。遗憾的是,将这两种方法结合起来不仅在技术上具有挑战性,而且随着线圈尺寸减小,其热指纹也会减小,从而降低了低温冷却的优势。在此,提出了一种替代解决方案,即在低温探头内采用伦兹透镜的形式。伦兹透镜并非取代检测线圈,而是能使来自较大线圈的B场重新聚焦到小得多的样品区域上。反过来,样品处更强的B场与低温线圈实现强耦合,从而改善信号。对于小样品,通过将内径为530的伦兹透镜与低温探头相结合,相对于单独使用低温探头,氢和碳的灵敏度分别进一步提高了2.8倍和3.5倍。此外,伦兹透镜的宽带特性使得能够研究多个原子核并采用异核二维(2D)NMR方法。在来自模式生物的430纳升血淋巴和八个卵(外径约350微米)上展示了灵敏度的提高和二维能力。总之,将伦兹透镜与低温探头相结合提供了一种相对简单的方法,可在保留低温探头优势的同时提高微小样品的灵敏度。