Cui Li, Yang Kai, Zhu Yong-Guan
Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen, China.
Methods Mol Biol. 2019;2046:95-107. doi: 10.1007/978-1-4939-9721-3_8.
Microorganisms play a key role in driving the global element (C, N, H, P, and S) cycling. However, the function and activity of environmental microbes remain largely elusive because the vast majority of them are yet uncultured. Recent achievements in single cell stable isotope-labeled Raman spectroscopy enable direct investigation of function and activity of individual microbes in complex environmental communities. Here, this protocol describes a workflow to investigate environmental microbes in soil and water by combining N, D, and C stable isotope labeling with different single-cell Raman techniques, including normal Raman, resonance Raman (RR), and surface-enhanced Raman spectroscopy (SERS). Their applications in investigating functional bacteria driving the N and C cycles, and metabolically active cells are described.
微生物在推动全球元素（碳、氮、氢、磷和硫）循环中发挥着关键作用。然而，环境微生物的功能和活性在很大程度上仍不为人所知，因为绝大多数微生物尚未得到培养。单细胞稳定同位素标记拉曼光谱技术的最新成果使得直接研究复杂环境群落中单个微生物的功能和活性成为可能。在此，本方案描述了一种通过将氮、氘和碳稳定同位素标记与不同的单细胞拉曼技术（包括普通拉曼、共振拉曼（RR）和表面增强拉曼光谱（SERS））相结合来研究土壤和水中环境微生物的工作流程。文中描述了它们在研究驱动氮和碳循环的功能细菌以及代谢活跃细胞方面的应用。
