Clark Amelia J, Masters-Clark Emily, Moratto Eleonora, Junier Pilar, Stanley Claire E
Department of Bioengineering, Imperial College London, London SW7 2AZ, United Kingdom.
Laboritoire de microbiologie, University of Neuchâtel, Neuchâtel CH-2000, Switzerland.
Biomicrofluidics. 2024 Oct 7;18(5):054109. doi: 10.1063/5.0231656. eCollection 2024 Sep.
Filamentous fungi and fungal-like organisms contribute to a wide range of important ecosystem functions. Evidence has shown the movement of liquid across mycelial networks in unsaturated environments, such as soil. However, tools to investigate liquid movement along hyphae at the level of the single cell are still lacking. Microfluidic devices permit the study of fungal and fungal-like organisms with cellular resolution as they can confine hyphae to a single optical plane, which is compatible with microscopy imaging over longer timescales and allows for precise control of the microchannel environment. The aim of this study was to develop a method that enables the visualization and quantification of liquid movement on hyphae of fungal and fungal-like microorganisms. For this, the fungal-fungal interaction microfluidic device was modified to allow for the maintenance of unsaturated microchannel conditions. Fluorescein-containing growth medium solidified with agar was used to track liquid transported by hyphae via fluorescence microscopy. Our key findings highlight the suitability of this novel methodology for the visualization of liquid movement by hyphae over varying time scales and the ability to quantify the movement of liquid along hyphae. Furthermore, we showed that at the cellular level, extracellular movement of liquid along hyphae can be bidirectional and highly dynamic, uncovering a possible link between liquid movement and hyphal growth characteristics. We envisage that this method can be applied to facilitate future research probing the parameters contributing to hyphal liquid movement and is an essential step for studying the phenomenon of fungal highways.
丝状真菌和类真菌生物对广泛的重要生态系统功能有贡献。有证据表明，在非饱和环境（如土壤）中，液体可在菌丝网络中移动。然而，目前仍缺乏在单细胞水平研究液体沿菌丝移动的工具。微流控装置能够以细胞分辨率研究真菌和类真菌生物，因为它们可以将菌丝限制在单个光学平面内，这与长时间的显微镜成像兼容，并能精确控制微通道环境。本研究的目的是开发一种方法，能够可视化和量化真菌及类真菌微生物菌丝上的液体移动。为此，对真菌 - 真菌相互作用微流控装置进行了改进，以维持非饱和微通道条件。用琼脂固化的含荧光素的生长培养基通过荧光显微镜来追踪菌丝运输的液体。我们的主要发现突出了这种新方法在不同时间尺度上可视化菌丝液体移动的适用性，以及量化液体沿菌丝移动的能力。此外，我们表明，在细胞水平上，液体沿菌丝的细胞外移动可以是双向的且高度动态的，揭示了液体移动与菌丝生长特性之间可能存在的联系。我们设想这种方法可用于促进未来对影响菌丝液体移动参数的研究探索，并且是研究真菌高速公路现象的重要一步。
