Fabrication and Use of the Dual-Flow-RootChip for the Imaging of Arabidopsis Roots in Asymmetric Microenvironments

Stanley, Claire E.; Shrivastava, Jagriti; Brugman, Rik; Heinzelmann, Elisa; Frajs, Viktoria; Bühler, Andreas; Swaay, Dirk van; Großmann, Guido

doi:10.21769/bioprotoc.3010

Cited by 10 publications

(6 citation statements)

References 17 publications

(6 reference statements)

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“…The design of the FFI device was constructed in AutoCAD Mechanical 2011 (Autodesk) and used to create a mylar® film photolithography mask (Micro Lithography Services Ltd., UK). The protocol for microfluidic device fabrication, including the manufacturing of the master mould, was previously described 19 , 49 . PDMS was formulated using a 10:1 ratio of base to curing agent (Sylgard 184, Dow Corning, USA) before degassing the mixture for 1 h under vacuum in a desiccator.…”

Section: Methodsmentioning

confidence: 99%

A versatile microfluidic platform measures hyphal interactions between Fusarium graminearum and Clonostachys rosea in real-time

et al. 2021

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Routinely, fungal–fungal interactions (FFI) are studied on agar surfaces. However, this format restricts high-resolution dynamic imaging. To gain experimental access to FFI at the hyphal level in real-time, we developed a microfluidic platform, a FFI device. This device utilises microchannel geometry to enhance the visibility of hyphal growth and provides control channels to allow comparisons between localised and systemic effects. We demonstrate its function by investigating the FFI between the biological control agent (BCA) Clonostachys rosea and the plant pathogen Fusarium graminearum. Microscope image analyses confirm the inhibitory effect of the necrotrophic BCA and we show that a loss of fluorescence in parasitised hyphae of GFP-tagged F. graminearum coincides with the detection of GFP in mycelium of C. rosea. The versatility of our device to operate under both water-saturated and nutrient-rich as well as dry and nutrient-deficient conditions, coupled with its spatio-temporal output, opens new opportunities to study relationships between fungi.

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Section: Methodsmentioning

confidence: 99%

A versatile microfluidic platform measures hyphal interactions between Fusarium graminearum and Clonostachys rosea in real-time

et al. 2021

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“…Microfluidic devices were prepared as described by Stanley et al based on a channel architecture that enables laminar flow conditions as a result of actively pumping solutions into the observation chamber (Figures and S3; cf. Supporting Information for detailed description).…”

Section: Methodsmentioning

confidence: 99%

“…Microfluidic Device Design and Preparation. Microfluidic devices were prepared as described by Stanley et al 47 based on a channel architecture 51 that enables laminar flow conditions as a result of actively pumping solutions into the observation chamber (Figures 1 and S3; cf. Supporting Information for detailed description).…”

Section: T H Imentioning

confidence: 99%

Mycelial Effects on Phage Retention during Transport in a Microfluidic Platform

Ghanem

Stanley

Harms

et al. 2019

Environ. Sci. Technol.

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Phages (i.e. viruses that infect bacteria) have been considered as good tracers for the hydrological transport of colloids and (pathogenic) viruses. Little, however, is known about interactions of phages with (fungal) mycelia as the prevalent soil microbial biomass. Forming extensive and dense networks, mycelia provide significant surfaces for phage-hyphal interactions. Here, we for the first time quantified the mycelial retention of phages in a microfluidic platform that allowed for defined fluid exchange around hyphae. Two common lytic tracer phages (Escherichia coli phage T4 and marine phage PSA-HS2) and two mycelia of differing surface properties (Coprionopsis cinerea, Pythium ultimum) were employed. Phage-hyphal interaction energies were approximated by the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) approach of colloidal interaction. Our data show initial hyphal retention of phages of up to ≈ 4 × 10 7 PFU mm-2 (≈ 2550 PFU mm-2 s-1) with a retention efficiency depending on the hyphal and, to a lesser extent, the phage surface properties. Experimental data were supported by XDLVO calculations, which revealed the highest attractive forces for the interaction between hydrophobic T4 phages and hydrophobic C. cinerea surfaces. Our data suggest that mycelia may be relevant for the retention of phages in the subsurface and need to be considered in subsurface phage tracer studies. Mycelia-phage interactions may further be exploited for the development of novel strategies to reduce or hinder the transport of undesirable (bio-)colloidal entities in environmental filter-systems.

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“…Microfluidic device fabrication and on-chip plant cultivation were performed as described in [88,89]. Briefly, Arabidopsis seedlings were grown in pipette tips filled with solidified medium and interfaced with poly(dimethylsiloxane) (PDMS) RootChip-8S devices 4-5 days after germination under sterile conditions.…”

Section: Microfluidic Device (Rootchip-8s) Fabrication and On-chip Plant Cultivationmentioning

confidence: 99%

Distinct RopGEFs Successively Drive Polarization and Outgrowth of Root Hairs

et al. 2019

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Fabrication and Use of the Dual-Flow-RootChip for the Imaging of Arabidopsis Roots in Asymmetric Microenvironments

Cited by 10 publications

References 17 publications

A versatile microfluidic platform measures hyphal interactions between Fusarium graminearum and Clonostachys rosea in real-time

A versatile microfluidic platform measures hyphal interactions between Fusarium graminearum and Clonostachys rosea in real-time

Mycelial Effects on Phage Retention during Transport in a Microfluidic Platform

Distinct RopGEFs Successively Drive Polarization and Outgrowth of Root Hairs

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