Environmental and Chemical Engineering Institute, Dalian University, Dalian, 116622, China; Dalian Key Laboratory of Oligosaccharide Recombination and Recombinant Protein Modification,Dalian, 116622, China.
Dalian Key Laboratory of Oligosaccharide Recombination and Recombinant Protein Modification,Dalian, 116622, China; Medical School, Dalian University, Dalian, 116622, China.
Anal Chim Acta. 2022 Sep 22;1227:340322. doi: 10.1016/j.aca.2022.340322. Epub 2022 Aug 28.
Microalgae are a group of photoautotrophic microorganisms which could use carbon dioxide for autosynthesis. They have been envisioned as one of the most prospective feedstock for renewable oil. However, great endeavors will still be needed to increase their economic feasibility. The screening of competitive species and suitable culture conditions are such issues. To greatly accelerate these rather laborious steps and also improve their experimental lump-sum-manner, we developed a microfluidic droplet-based 2 × 10 resolution "identification card", which allowed high throughput real-time monitoring of individual algae among population. A novel fluid-blocking-based droplet generating and trapping performance were integrated in the platform which made it excellent in operational simplicity, rapidity and stability and full of the potentials in single-cell-isolation/screening. The developed platform was successfully used to screen three unicellular algae, namely, Isochrysis zhanjiangensis, Platymonas subcordiformis and Platymonas helgolandica var. tsingtaoensis. In situ bioassays of the lipid accumulation and cell proliferation at single cell level for interspecies comparison were possible. Furthermore, lipid-producing inhomogeneity was demonstrated among cells in the same specie and batch. Nitrogen stress condition can be identified that induce positive-skewed frequency distribution of lipid content, even among individual inhomogeneous cells over the typically used culture condition.
微藻是一群光合自养微生物，能够利用二氧化碳进行自养合成。它们被认为是最有前途的可再生油原料之一。然而，为了提高其经济可行性，仍需要做出巨大的努力。竞争物种的筛选和适宜的培养条件就是此类问题。为了大大加快这些相当繁琐的步骤，并提高其实验的整体效率，我们开发了一种基于微流控的 2×10 分辨率的“识别卡”，它可以实现对群体中单个藻类的高通量实时监测。该平台集成了一种基于新型流体阻塞的液滴生成和捕获性能，使其在操作简单性、快速性和稳定性方面表现出色，并且在单细胞分离/筛选方面具有很大的潜力。所开发的平台成功地用于筛选三种单细胞藻类，即湛江等鞭金藻、亚心形扁藻和青岛大扁藻。可以在单细胞水平上对不同物种的脂积累和细胞增殖进行原位生物测定。此外，还证明了同一物种和批次的细胞之间存在产脂不均匀性。可以识别氮胁迫条件，其导致脂质含量的正偏态频率分布，即使在典型培养条件下的不均匀细胞之间也是如此。
