Soil imaging in the field and laboratory has greatly advanced our understanding of plant root systems. Soil fungi function as important plant symbionts and decomposers of complex organic material in soil environments. For fungal hyphae, however, the application of soil imaging remains scarce, limiting our understanding of hyphal systems in soil. This scarce application is partly due to the challenging development of a soil imaging device for hyphae: technical requirements to resolve fine hyphae (2-5 μm in diameter) are high, while the device cost must be low to facilitate sufficient deployment that can capture the high spatial heterogeneity of hyphal dynamics in soil. This protocol describes the do-it-yourself assembly and application of a low-cost high-resolution imaging device for observing hyphae in soil. The assembly of the open-source imaging device relies on many 3D-printed parts, reducing material costs to ca. 930 USD. The application of the imaging device yields soil profile images with a resolution of up to 0.52 μm px-1 (49000 dpi) within an observable volume of 70 × 210 × 1.5 mm. By repeatedly imaging a soil profile using the presented techniques, changes in the amount, distribution, and morphology of hyphae in soil can be observed and quantified.
Copyright: © 2025 Holger Schaefer. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.