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微藻生物技术过程开发中的流式细胞术。

Flow cytometry for the development of biotechnological processes with microalgae.

机构信息

Institute of Biotechnology, Zurich University of Applied Sciences (ZHAW), Campus Grüental, CH-8820 Wädenswil, Switzerland.

出版信息

Biotechnol Adv. 2013 Jan-Feb;31(1):2-16. doi: 10.1016/j.biotechadv.2012.04.007. Epub 2012 Apr 25.

Abstract

The current interest in microalgae as a sustainable source of next generation biofuels and other valuable substances is driving exploration of their use as unique biotechnological production systems. To design and optimise appropriate production strategies, the behaviour of particular microalgal species should be well characterised under different culture conditions. Thus, flow cytometric (FCM) methods, which are already well established in environmental and toxicological studies of microalgae, are also useful for analysing the physiological state of microalgae, and have the potential to contribute to the rapid development of feasible bioprocesses. These methods are commonly based on the examination of intrinsic features of individual cells within a population (such as autofluorescence or size). Cells possessing the desired physiological or morphological features, which are detectable with or without fluorescent staining, are counted or isolated (sorted) using an FCM device. The options for implementation of FCM in the development of biotechnological processes detailed in this review are (i) analysing the chemical composition of biomass, (ii) monitoring cellular enzyme activity and cell viability, and (iii) sorting cells to isolate those overproducing the target compound or for the preparation of axenic cultures.

摘要

当前,人们对微藻作为下一代生物燃料和其他有价值物质的可持续来源的兴趣日益浓厚,这促使人们探索将其用作独特的生物技术生产系统。为了设计和优化合适的生产策略,应在不同的培养条件下对特定微藻物种的行为进行充分表征。因此,流式细胞术(FCM)方法在微藻的环境和毒理学研究中已经得到很好的应用,也可用于分析微藻的生理状态,并有可能有助于快速开发可行的生物工艺。这些方法通常基于对群体内单个细胞的固有特征(如自发荧光或大小)的检查。使用 FCM 设备,可以对具有所需生理或形态特征的细胞进行计数或分离(分选),这些特征可以通过荧光染色或不进行荧光染色来检测。本综述详细介绍了在生物技术工艺开发中实施 FCM 的几种选择,包括:(i)分析生物量的化学成分,(ii)监测细胞酶活性和细胞活力,以及(iii)对细胞进行分选以分离目标化合物过量产生的细胞或用于制备无菌培养物。

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