Microfluidic Sensors for the Detection of Motile Plant Zoospores

24 September 2024, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Plant pathogen zoospores play a vital role in the transmission of several significant plant diseases, with their early detection being important for effective pathogen management. Current methods for pathogen detection involve labour-intensive specimen collection and laboratory testing, lacking real-time feedback capabilities. Methods that can be deployed in the field and remotely addressed are required. In this proof-of-concept study, we have developed an innovative zoospore-sensing device by combining a microfluidic system that interfaced a flow stream with a microfluidic cytometer, whilst incorporating a chemotactic response as a means to selectively detect motile spores. Spores of Phytophthora cactorum were captured in eddies in a stub at a right-angle bend in a flow channel from which they were able to swim up a detection channel following a gradient of attractant. They were then detected by a transient change in impedance, as measured with a single-chip lock-in amplifier, when they passed between a pair of electrodes. Such a sensing system has a great potential to be further developed into a portable, remotely addressable, low-cost sensing system, offering an important tool for field pathogen real-time detection applications.

Keywords

zoospores
microfluidics
cytometer
chemotaxis
plant pathogen detection
field instrumentation

Supplementary materials

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supplementary information
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1. Microfluidic chip fabrication 2. Phytophthora cactorum zoospore production 3. Chemotaxis of zoospores 4. Electronic design and recording set-up 5. Simulation results for various channel heights and electrode spacings 6. Phase separation during the channel filling Video 1: Release of Phytophthora cactorum zoospores Video 2: Chemotaxis of Phytophthora cactorum zoospores Video 3: Zoospore sensing in the pumping setup Video 4: Zoospore sensing with them swimming through the channel under the gradient of attractant
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Video 1: Release of Phytophthora cactorum zoospores
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Video microscopy showing the release of motile spores
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Video 2: Chemotaxis of Phytophthora cactorum zoospores
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Video microscopy showing spores swimming up concentration gradient of attractant
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Video 3: Zoospore sensing in the pumping setup
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Video microscopy showing spores translating along the channel in a pumped flow, illustrating correlation of zoospore passage between the electrodes with an electrical signal
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Video 4: Zoospore sensing with them swimming through the channel under the gradient of attractant
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Video microscopy showing motile spores swimming along the microfluidic channel, up a concentration gradient of attractant, with passage between the electrodes detected by the correlated electrical signal.
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