Single-cell Center, CAS Key Laboratory of Biofuels, and Shandong Key Laboratory of Energy Genetics, Qingdao Institute of BioEnergy and Bioprocess Technology, Chinese Academy of Sciences , Qingdao, Shandong, 266101, P. R. China.
Anal Chem. 2013 Nov 19;85(22):10697-701. doi: 10.1021/ac403107p. Epub 2013 Oct 28.
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 (13)C and (15)N 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).
我们优化了一台拉曼显微镜，以在 0.1 秒的采集时间内获得单个细胞拉曼光谱（SCRS）。具有如此短采集时间的 SCRS 具有足够的区分能力和光谱重现性，可区分掺入（13）C 和（15）N 的细胞，并对从口腔中分离出的五种不同类型的细菌进行分类。我们还开发了拉曼激活细胞弹射（RACE），该技术得到了激光诱导正向转移（LIFT）的辅助。我们首次表明，可以根据 SCRS 从复杂的微生物群落中识别和准确分离出感兴趣的单个细胞。这种方法可用于从复杂样品（例如生物膜、土壤、污泥、组织）中对具有目标特征的单个细胞进行分选。
