Rao Wei, Pan Ning, Yang Zhibo
Department of Chemistry and Biochemistry, University of Oklahoma.
Department of Chemistry and Biochemistry, University of Oklahoma;
J Vis Exp. 2016 Jun 14(112):53911. doi: 10.3791/53911.
Mass spectrometry imaging (MSI) and in-situ single cell mass spectrometry (SCMS) analysis under ambient conditions are two emerging fields with great potential for the detailed mass spectrometry (MS) analysis of biomolecules from biological samples. The single-probe, a miniaturized device with integrated sampling and ionization capabilities, is capable of performing both ambient MSI and in-situ SCMS analysis. For ambient MSI, the single-probe uses surface micro-extraction to continually conduct MS analysis of the sample, and this technique allows the creation of MS images with high spatial resolution (8.5 µm) from biological samples such as mouse brain and kidney sections. Ambient MSI has the advantage that little to no sample preparation is needed before the analysis, which reduces the amount of potential artifacts present in data acquisition and allows a more representative analysis of the sample to be acquired. For in-situ SCMS, the single-probe tip can be directly inserted into live eukaryotic cells such as HeLa cells, due to the small sampling tip size (< 10 µm), and this technique is capable of detecting a wide range of metabolites inside individual cells at near real-time. SCMS enables a greater sensitivity and accuracy of chemical information to be acquired at the single cell level, which could improve our understanding of biological processes at a more fundamental level than previously possible. The single-probe device can be potentially coupled with a variety of mass spectrometers for broad ranges of MSI and SCMS studies.
常压条件下的质谱成像(MSI)和原位单细胞质谱(SCMS)分析是两个新兴领域,在对生物样品中的生物分子进行详细质谱(MS)分析方面具有巨大潜力。单探针是一种具有集成采样和电离功能的小型化设备,能够同时进行常压MSI和原位SCMS分析。对于常压MSI,单探针使用表面微萃取对样品持续进行MS分析,该技术能够从诸如小鼠脑和肾切片等生物样品中创建具有高空间分辨率(8.5 µm)的MS图像。常压MSI的优势在于分析前几乎无需样品制备,这减少了数据采集中潜在伪像的数量,并能获取更具代表性的样品分析结果。对于原位SCMS,由于采样尖端尺寸小(< 10 µm),单探针尖端可直接插入活的真核细胞,如HeLa细胞,并且该技术能够近乎实时地检测单个细胞内的多种代谢物。SCMS能够在单细胞水平上获取更高灵敏度和准确性的化学信息,这比以往任何时候都能在更基础的层面上增进我们对生物过程的理解。单探针设备有可能与各种质谱仪联用,用于广泛的MSI和SCMS研究。