The need for speed: Stimulated Raman Spectroscopy for human gut microbiome research

我要求对我的R35 Mira奖进行补充，以帮助获得用于受激拉曼的激光光源 用显微镜帮助我的实验室研究开发用于人体肠道微生物组的新型显微镜工具 研究。在我最初的Mira应用程序中，为人类开发下一代生理学方法 肠道微生物组研究(奖1R35GM147166-01)，我描述了强大但目前未得到充分利用的 拉曼显微光谱学在人体肠道微生物组研究中的潜力。具体来说，我 阐述了自发拉曼显微镜将如何用于研究方向2和3，这将 结合稳定同位素探测和拉曼光谱探测微生物原位功能(方向2)和关联 拉曼与其他显微技术相结合，以更全面地了解肠道微生物(方向 3)。受激拉曼光谱的采集速度比自发采集速度快1000-100万 拉曼光谱，但直到最近才能在商业上获得；当徕卡推出 第一台上市的自发拉曼显微镜。这台仪器的能力将允许光谱 采购将增加1,000倍，这将使结肠镜检查和其他人类肠道检查成为可能 微生物样本将在几分钟内以亚细胞空间分辨率进行化学测绘。此外, 这将使我的实验室扩大我们的研究，以可视化微生物细胞身份和合成代谢活动使用 振动探针，这是目前使用自发拉曼技术所不可能的，因此没有 包括在我最初的Mira应用程序中。如果获奖，该乐器将成为第三个被激发的乐器 拉曼显微镜在全国任何一所大学都有。

I request a supplement to my R35 MIRA award to help acquire the laser source for a Stimulated Raman Microscope to aid my lab’s research on developing new microscopy tools for human gut microbiome research. In my original MIRA application, Developing Next-Generation Physiology approaches for human gut microbiome research (award 1R35GM147166-01), I described the strong but currently underutilized potential of Raman microspectroscopy (Raman) for human gut microbiome research. Specifically, I addressed how Spontaneous Raman microscopy will be used in research directions 2 and 3, which will combine stable isotope probing and Raman to probe microbial in situ function (direction 2) and correlate Raman with other microscopic techniques to gain a more holistic understanding of gut microbes (direction 3). Stimulated Raman spectroscopy offers 1,000-1,000,000 faster acquisition rates than Spontaneous Raman spectroscopy, but until recently was not commercially available; this changed when Leica brought the first Spontaneous Raman microscope to market. The capabilities of this instrument will allow spectral acquisition to be increased by a factor of 1,000, which will enable colonoscopy and other human gut microbiome samples to be chemically mapped at subcellular spatial resolution within minutes. In addition, it will allow my lab to expand our research to visualize microbial cell identity and anabolic activity using vibrational probes, which is not possible using currently Spontaneous Raman technology and thus was not included in my original MIRA application. If awarded, this instrument will become the 3rd Stimulated Raman microscope at any university in the nation.