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首页> 外文期刊>Applied Microbiology and Biotechnology >Morphological analysis of the filamentous fungus Penicillium chrysogenum using flow cytometry-the fast alternative to microscopic image analysis
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Morphological analysis of the filamentous fungus Penicillium chrysogenum using flow cytometry-the fast alternative to microscopic image analysis

机译：流式细胞术的丝状真菌青霉甲烯源的形态学分析 - 微观图像分析的快速替代品

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摘要

An important parameter in filamentous bioreactor cultivations is the morphology of the fungi, due to its interlink to productivity and its dependency on process conditions. Filamentous fungi show a large variety of morphological forms in submerged cultures. These range from dispersed hyphae, to interwoven mycelial aggregates, to denser hyphal aggregates, the so-called pellets. Depending on the objective function of the bioprocess, different characteristics of the morphology are favorable and need to be quantified accurately. The most common method to quantitatively characterize morphology is image analysis based on microscopy. This method is work intensive and time consuming. Therefore, we developed a faster, at-line applicable, alternative method based on flow cytometry. Within this contribution, this novel method is compared to microscopy for a penicillin production process. Both methods yielded in comparable distinction of morphological sub-populations and described their morphology in more detail. In addition to the appropriate quantification of size parameters and the description of the hyphal region around pellets, the flow cytometry method even revealed a novel compactness parameter for fungal pellets which is not accessible via light microscopy. Hence, the here presented flow cytometry method for morphological analysis is a fast and reliable alternative to common tools with some new insights in the pellet morphology, enabling at-line use in production environments.
机译：丝状生物反应器培养中的一个重要参数是真菌的形态，因为它对生产力的交互及其对工艺条件的依赖性。丝状真菌在淹没培养物中显示出各种形态形态。这些范围从分散的菌丝，交织菌丝体聚集体，以更密集的悬垂聚集体，所谓的颗粒。根据生物过程的目标函数，形态的不同特征是有利的，需要准确地量化。定量表征形态学的最常见方法是基于显微镜的图像分析。这种方法是强化和耗时的工作。因此，我们开发了一种基于流式细胞术的更快，齐全的适用，替代方法。在此贡献中，将这种新方法与Penicillin生产过程的显微镜进行比较。两种方法在形态亚群的相当区别中产生，并更详细地描述了它们的形态。除了尺寸参数的适当定量和沉淀周围的亚腿区域的描述之外，流式细胞术方法甚至揭示了对真菌颗粒的新型紧凑性参数，其不通过光学显微镜接近。因此，这里提出了用于形态学分析的流式细胞术方法是一种快速可靠的替代常见工具，具有一些新的颗粒形态的新见解，在生产环境中可以在线使用。

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