以细胞分辨率观察液体在整个菌丝网络中的分布。

IF 2.6 4区 工程技术 Q2 BIOCHEMICAL RESEARCH METHODS
Biomicrofluidics Pub Date : 2024-10-07 eCollection Date: 2024-09-01 DOI:10.1063/5.0231656
Amelia J Clark, Emily Masters-Clark, Eleonora Moratto, Pilar Junier, Claire E Stanley
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引用次数: 0

摘要

丝状真菌和类真菌生物对生态系统的多种重要功能做出了贡献。有证据表明,在土壤等非饱和环境中,液体会通过菌丝网络流动。然而,在单细胞水平上研究液体沿菌丝运动的工具仍然缺乏。微流体设备可以将菌丝限制在单个光学平面内,因此可以对真菌和类真菌生物进行细胞分辨率的研究,这与较长时间尺度的显微镜成像兼容,并可对微通道环境进行精确控制。本研究的目的是开发一种方法,使真菌和类真菌微生物菌丝上的液体运动可视化和定量化。为此,对真菌-真菌相互作用微流控装置进行了改进,使其能够维持不饱和微通道条件。含荧光素的生长培养基用琼脂固化,通过荧光显微镜追踪菌丝输送的液体。我们的主要研究结果突出表明,这种新方法适用于在不同时间尺度上观察液体通过菌丝的运动,并能量化液体沿菌丝的运动。此外,我们还发现,在细胞水平上,细胞外液体沿菌丝的运动可以是双向的,而且是高度动态的,这揭示了液体运动与菌丝生长特性之间可能存在的联系。我们认为,这种方法可用于促进未来的研究,探究导致菌丝液体运动的参数,是研究真菌高速公路现象的重要一步。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Visualizing liquid distribution across hyphal networks with cellular resolution.

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.

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来源期刊
Biomicrofluidics
Biomicrofluidics 生物-纳米科技
CiteScore
5.80
自引率
3.10%
发文量
68
审稿时长
1.3 months
期刊介绍: Biomicrofluidics (BMF) is an online-only journal published by AIP Publishing to rapidly disseminate research in fundamental physicochemical mechanisms associated with microfluidic and nanofluidic phenomena. BMF also publishes research in unique microfluidic and nanofluidic techniques for diagnostic, medical, biological, pharmaceutical, environmental, and chemical applications. BMF offers quick publication, multimedia capability, and worldwide circulation among academic, national, and industrial laboratories. With a primary focus on high-quality original research articles, BMF also organizes special sections that help explain and define specific challenges unique to the interdisciplinary field of biomicrofluidics. Microfluidic and nanofluidic actuation (electrokinetics, acoustofluidics, optofluidics, capillary) Liquid Biopsy (microRNA profiling, circulating tumor cell isolation, exosome isolation, circulating tumor DNA quantification) Cell sorting, manipulation, and transfection (di/electrophoresis, magnetic beads, optical traps, electroporation) Molecular Separation and Concentration (isotachophoresis, concentration polarization, di/electrophoresis, magnetic beads, nanoparticles) Cell culture and analysis(single cell assays, stimuli response, stem cell transfection) Genomic and proteomic analysis (rapid gene sequencing, DNA/protein/carbohydrate arrays) Biosensors (immuno-assay, nucleic acid fluorescent assay, colorimetric assay, enzyme amplification, plasmonic and Raman nano-reporter, molecular beacon, FRET, aptamer, nanopore, optical fibers) Biophysical transport and characterization (DNA, single protein, ion channel and membrane dynamics, cell motility and communication mechanisms, electrophysiology, patch clamping). Etc...
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