青岛生物能源与过程研究所高通量单细胞拉曼分选技术取得新进

现有基于荧光染色的活体单细胞分选技术(Fluorescence-Activated Cell Sorting; FACS)一般需要外加荧光标记，在单位时间只能获得个别分子所代表的表型，且通常难以直接进行“原位”观测。而基于拉曼显微光谱技术的单细胞分选方法无需外加标记，可无损获得整个单细胞的化学物质指纹图谱，从而迅速识别活体单细胞的种系发生、生理特性和代谢产物变化等，因此对于难培养微生物的功能鉴定和资源开发具有重要意义。

近日，中国科学院青岛生物能源与过程研究所单细胞研究中心功能基因组团队助理研究员王允、籍月彤等通过优化光路、提高激光强度等方式显著提高了单细胞拉曼光谱的采集速度，首次将微生物细胞拉曼图谱获取时间降低到毫秒级，为高通量单细胞拉曼分选奠定了基础。同时，结合激光诱导向前转移(LIFT)原理，发明了拉曼激活细胞弹射(Raman-activated Cell Ejection;RACE)技术，可用于将特定拉曼表型的单细胞从复杂微生物群落中分离，从而获取其基因组信息。这一工作为单细胞拉曼分选仪(Raman-activated Cell Sorter;RACS)提供了新的一种分选方法。相关研究成果发表于最新一期的Analytical Chemistry。

上述研究由英国Sheffield大学黄巍教授和青岛能源所单细胞研究中心主任徐健研究员合作主持完成。

Raman Activated Cell Ejection for Isolation of Single Cells

Yun Wang, Yuetong Ji, Emma S. Wharfe, Roger S. Meadows, Peter March, Royston Goodacre
, Jian Xu , and Wei E. Huang

We have optimized a Raman microscope to obtain a single cell Raman spectrum (SCRS) with 0.1 s acquisition time. SCRS with such short acquisition time has sufficient discriminatory ability and spectral reproducibility to differentiate cells incorporated with 13C and 15N and to classify five different types of bacteria isolated from the oral cavity. We also developed Raman activated cell ejection (RACE) that is assisted by laser induced forward transfer (LIFT). We have shown, for the first time, that the single cells of interest can be identified and then accurately isolated from complex microbial communities based on their SCRS. This approach can be used to sort single cells of target traits from complex samples (e.g., biofilms, soils, sludge, tissues).