Zhang Qiang, Zhang Peiran, Gou Honglei, Mou Chunbo, Huang Wei E, Yang Menglong, Xu Jian, Ma Bo
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, China.
Analyst. 2015 Sep 21;140(18):6163-74. doi: 10.1039/c5an01074h.
Raman-activated cell sorting (RACS) is a promising single-cell analysis technology that is able to identify and isolate individual cells of targeted type, state or environment from an isogenic population or complex consortium of cells, in a label-free and non-invasive manner. However, compared with those widely used yet labeling-required or staining-dependent cell sorting technologies such as FACS and MACS, the weak Raman signal greatly limits the further development of the existing RACS systems to achieve higher throughput. Strategies that can tackle this bottleneck include, first, improvement of Raman-acquisition efficiency and quality based on advanced Raman spectrometers and enhanced Raman techniques; second, development of novel microfluidic devices for cell sorting followed by integration into a complete RACS system. Exploiting these strategies, prototypes for a new generation of RACS have been demonstrated, such as flow-based OT-RACS, DEP-RACS, and SERS/CARS flow cytometry. Such high-throughput microfluidic RACS can provide biologists with a powerful single-cell analysis tool to explore the scientific questions or applications that have been beyond the reach of FACS and MACS.
拉曼激活细胞分选（RACS）是一项很有前景的单细胞分析技术，它能够以无标记、非侵入的方式，从同基因细胞群体或复杂细胞聚集体中识别并分离出具有特定类型、状态或环境的单个细胞。然而，与那些广泛使用但需要标记或依赖染色的细胞分选技术（如荧光激活细胞分选术（FACS）和磁性激活细胞分选术（MACS））相比，微弱的拉曼信号极大地限制了现有RACS系统实现更高通量的进一步发展。解决这一瓶颈的策略包括：第一，基于先进的拉曼光谱仪和增强拉曼技术提高拉曼采集效率和质量；第二，开发用于细胞分选的新型微流控装置，然后将其集成到完整的RACS系统中。利用这些策略，新一代RACS的原型已经得到展示，如基于流动的光镊拉曼激活细胞分选（OT-RACS）、介电电泳拉曼激活细胞分选（DEP-RACS）以及表面增强拉曼散射/相干反斯托克斯拉曼散射流式细胞术（SERS/CARS流式细胞术）。这种高通量微流控RACS能够为生物学家提供一个强大的单细胞分析工具，以探索那些FACS和MACS无法触及的科学问题或应用。
