Yu Ziyi, Geisler Katrin, Leontidou Tina, Young Rosanna E B, Vonlanthen Sofie E, Purton Saul, Abell Chris, Smith Alison G
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, UK.
Algal Res. 2021 Jun;56:None. doi: 10.1016/j.algal.2021.102293.
The application of microfluidic technologies to microalgal research is particularly appealing since these approaches allow the precise control of the extracellular environment and offer a high-throughput approach to studying dynamic cellular processes. To expand the portfolio of applications, here we present a droplet-based microfluidic method for analysis and screening of and , which can be integrated into a genetic transformation workflow. Following encapsulation of single cells in picolitre-sized droplets, fluorescence signals arising from each cell can be used to assess its phenotypic state. In this work, the chlorophyll fluorescence intensity of each cell was quantified and used to identify populations of cells grown in different light conditions. Further, individual or cells engineered to express green fluorescent protein were distinguished and sorted from wild-type cells. This has been exploited as a rapid screen for transformed cells within a population, bypassing a major bottleneck in algal transformation workflows and offering an alternative strategy for the identification of genetically modified strains.
微流控技术在微藻研究中的应用特别具有吸引力，因为这些方法能够精确控制细胞外环境，并为研究动态细胞过程提供高通量方法。为了扩展应用范围，我们在此展示一种基于液滴的微流控方法，用于分析和筛选[具体内容缺失]，该方法可整合到基因转化工作流程中。在将单细胞封装到皮升大小的液滴中后，每个细胞产生的荧光信号可用于评估其表型状态。在这项工作中，对每个细胞的叶绿素荧光强度进行了量化，并用于识别在不同光照条件下生长的[具体内容缺失]细胞群体。此外，对经过基因改造以表达绿色荧光蛋白的单个[具体内容缺失]或[具体内容缺失]细胞与野生型细胞进行了区分和分选。这已被用作快速筛选群体中转化细胞的方法，绕过了藻类转化工作流程中的一个主要瓶颈，并为鉴定转基因菌株提供了一种替代策略。
