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通过介电泳法分离异养微藻

Separation of Heterotrophic Microalgae by Dielectrophoresis.

作者信息

Birkholz Mario, Malti Danai Eleni, Hartmann Stephan, Neubauer Peter

机构信息

IHP-Leibniz-Institut für Innovative Mikroelektronik, Frankfurt, Germany.

Department of Biotechnology, Chair of Bioprocess Engineering, Technische Universität Berlin, Berlin, Germany.

出版信息

Front Bioeng Biotechnol. 2022 May 23;10:855035. doi: 10.3389/fbioe.2022.855035. eCollection 2022.

Abstract

Microalgae constitute an abundant source of poly-unsaturated fatty acids which are applied in various biotechnological fields such as pharmaceuticals and food supplement. Separating microalgae cells with respect to their lipid content would establish a relevant analytical technique. The present study demonstrates an electrical approach for the separation of the lipid-producing microalgae using the effect of dielectrophoresis (DEP) in a microfluidic flow cell. Microalgae were cultivated for 8 days, while cell growth was characterized by optical density, dry cell weight, glucose concentration and lipid content fluorescence microscopy. The size distribution of cells during cultivation was thoroughly investigated, since the DEP force scales with cell volume, but also depends on lipid content cell electrophysiological constants. Thus, the challenge was to deconvolute one separation effect from the other, while the electrical cell constants of are not known yet. The DEP-dependent separation was realized by slanted top-bottom electrodes with the flowing cell suspension between them. Turning on the voltage deflected the cells from their initial path as determined by the streaming and thus changed their direction of flow. The separation efficiency of DEP was tested for various electrical field strengths and its performance was determined by quantitative analysis of optical and fluorescence videos. It could be shown for all size groups that the most lipid-containing cells were always subject to DEP separation and that the method is thus not only suitable for process analysis, but also for strain selection of the most productive cell lines.

摘要

微藻是多不饱和脂肪酸的丰富来源,这些脂肪酸应用于制药和食品补充剂等各种生物技术领域。根据脂质含量分离微藻细胞将建立一种相关的分析技术。本研究展示了一种在微流控流动池中利用介电泳(DEP)效应分离产脂微藻的电学方法。微藻培养8天,通过光密度、干细胞重量、葡萄糖浓度和脂质含量荧光显微镜对细胞生长进行表征。对培养过程中细胞的大小分布进行了深入研究,因为DEP力与细胞体积有关,但也取决于脂质含量和细胞电生理常数。因此,挑战在于将一种分离效应与另一种分离效应解卷积,而细胞的电学常数尚不清楚。通过倾斜的上下电极实现DEP依赖的分离,流动的细胞悬浮液位于电极之间。开启电压会使细胞偏离由流动决定的初始路径,从而改变其流动方向。针对不同电场强度测试了DEP的分离效率,并通过对光学和荧光视频的定量分析来确定其性能。对于所有大小组都可以表明,脂质含量最高的细胞总是会受到DEP分离,因此该方法不仅适用于过程分析,也适用于选择最高产细胞系的菌株。

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