基于液滴-质谱联用的单细胞精度微生物组代谢分析新方法研究

代谢物组是解析微生物组功能机制的重要工具。但是微生物组通常结构复杂，其个体成员的代谢组一般难以从传统意义上的元代谢物组数据中得以重建，这是当前元代谢组实验策略的核心局限性之一。针对这一瓶颈，本项目拟采用微液滴分散包裹微生物，即将单个(或数个共生)微生物细胞包裹在微液滴中进行高通量并行培养，并且与质谱耦合进行代谢组学分析，力图实现在“个体”层面全面解析微生物组代谢谱。拟重点建立液滴单细胞培养及分选、芯片-MALDI自动点样和在线原位代谢物预处理等关键技术，进而以口腔微生物组为模式检测体系，探索一种以单细胞克隆化群体为并行分析单元、超高速、一体化的微生物组代谢分析新方法。

Meta-metabolomics is an important tool for dissecting the functional mechanisms of microbiota. However, due to the extraordinary complexity of microbiota, reconstructing the metabolome of individual members of a microbiota from traditional meta-metabolomics datasets is typically quite difficult. This has become one key limitation of the current experimental strategy of meta-metabolomics. To tackle this problem, this proposal aims to employ microfluidic droplets, in which single (or several symbiotic) microbial cells are wrapped in micro-droplets for high-throughput parallel culture. The droplet-based single-cell culture is then coupled to downstream mass spectrometry for metabolomics analysis, in parallel. To achieve this goal, several key techniques will be developed, which include: single-cell cultivation in droplets, auto-spotting of matrix-assisted laser desorption/ionization (MALDI) interface, and online in situ pretreatment of samples. In the end, an innovative single-cell precision (i.e., from clonal population of single-cells) ultra-high-speed integrated metabolomics method would be demonstrated, using oral microbiota as an example.