Large-scale label-free single-cell analysis of paramylon in Euglena gracilis by high-throughput broadband Raman flow cytometry
Large-scale label-free single-cell analysis of paramylon in Euglena gracilis by high-throughput broadband Raman flow cytometry
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DOI：
10.1364/boe.382957
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发表时间：
2020-04-01
影响因子：
3.4
通讯作者：
Goda, Keisuke
中科院分区：
文献类型：
--
作者：
Hiramatsu, Korato;Yamada, Koji;Goda, Keisuke
基于微藻的生物材料生产作为药物、食品和生物燃料的新来源已受到关注。为了提高生产效率，了解异质微藻亚群之间的差异至关重要。然而，由于缺乏单细胞分辨率或无法进行大规模分析以及检测小分子，现有技术不足以满足需求。在此，我们利用我们最近开发的高通量宽带拉曼流式细胞仪，以>1000个细胞/秒的通量，对小眼虫（一种单细胞微藻物种，其产生副淀粉作为治疗艾滋病和结肠癌的潜在药物）进行了大规模单细胞分析。具体而言，我们根据在五种不同条件下培养的10,000个小眼虫细胞的单细胞拉曼光谱表征了副淀粉的细胞内含量，并发现在相同条件下培养的小眼虫细胞中的副淀粉含量呈对数正态分布，这是描述反应网络中化学物质数量的一个良好模型。以无标记方式表征分布函数的能力是基于代谢物的细胞内含量通过定向进化分离用于合成生物学的特定细胞群的重要基础。（C）2020美国光学学会，依据美国光学学会开放获取出版协议条款
Microalga-based biomaterial production has attracted attention as a new source of drugs, foods, and biofuels. For enhancing the production efficiency, it is essential to understand its differences between heterogeneous microalgal subpopulations. However, existing techniques are not adequate to address the need due to the lack of single-cell resolution or the inability to perform large-scale analysis and detect small molecules. Here we demonstrated large-scale single-cell analysis of Euglena gracilis (a unicellular microalgal species that produces paramylon as a potential drug for HIV and colon cancer) with our recently developed high-throughput broadband Raman flow cytometer at a throughput of >1,000 cells/s. Specifically, we characterized the intracellular content of paramylon from single-cell Raman spectra of 10,000 E. gracilis cells cultured under five different conditions and found that paramylon contents in E. gracilis cells cultured in an identical condition is given by a log-normal distribution, which is a good model for describing the number of chemicals in a reaction network. The capability of characterizing distribution functions in a label-free manner is an important basis for isolating specific cell populations for synthetic biology via directed evolution based on the intracellular content of metabolites. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement