Highly Efficient Capture and Quantification of the Airborne Fungal Pathogen Sclerotinia sclerotiorum Employing a Nanoelectrode-Activated Microwell Array

Abstract

In this study, we present a microdevice for the capture and quantification of Sclerotinia sclerotiorum spores, pathogenic agents of one of the most harmful infectious diseases of crops, Sclerotinia stem rot. The early prognosis of an outbreak is critical to avoid severe economic losses and can be achieved by the detection of a small number of airborne spores. However, the current lack of simple and effective methods to quantify fungal airborne pathogens has hindered the development of an accurate early warning system. We developed a device that remedies these limitations based on a microfluidic design that contains a nanothick aluminum electrode structure integrated with a picoliter well array for dielectrophoresis-driven capture of spores and on-chip quantitative detection employing impedimetric sensing. Based on experimental results, we demonstrated a highly efficient spore trapping rate of more than 90% with an effective impedimetric sensing method that allowed the spore quantification of each column in the array and achieved a sensitivity of 2%/spore at 5 kHz and 1.6%/spore at 20 kHz, enabling single spore detection. We envision that our device will contribute to the development of a low-cost microfluidic platform that could be integrated into an infectious plant disease forecasting tool for crop protection.

Highly efficient cell-microbead encapsulation using dielectrophoresis-assisted dual-nanowell array

“…The ability to be easily upscaled suggests that our device may be comparable with commercial products that typically handle tens of thousands of cells. Since DEP force depends on the dielectric property of cells, the selective capture of a target cell type from a mixed sample with this label-free approach is possible ( 31 , 34 , 48 ). Our microfluidic device is expected to integrate cell sorting and analysis functions into one chip, which is promising for many applications, including the analysis of CTCs ( 21 ), immunocytes ( 49 ), and cell–cell complexes ( 50 ) from peripheral blood samples.…”

Microfluidics in environmental analysis: advancements, challenges, and future prospects for rapid and efficient monitoring

“…The device achieves over 90% spore trapping rate and high sensitivity, enabling single spore detection, making it valuable for crop disease forecasting. 277…”

Bionanotechnology and bioMEMS (BNM): state-of-the-art applications, opportunities, and challenges

“…The ability to probe and discriminate based on differences in the cytoplasm is vital where cells may be broadly similar, such as discriminating cancer cells from healthy ones, 154–157 identifying stem cell differentiation fate, 158–160 or trapping spores that can affect agriculture food plants. 161,162…”

“…The ability to probe and discriminate based on differences in the cytoplasm is vital where cells may be broadly similar, such as discriminating cancer cells from healthy ones, [154][155][156][157] identifying stem cell differentiation fate, [158][159][160] or trapping spores that can affect agriculture food plants. 161,162 A classical electrostatic method for microfluidic bioparticle concentration, separation and analysis is dielectrophoresis (DEP). 163 In DEP, an electric field is used to induce a dipole in a bioparticle of interest, and a gradient is applied which creates a net force.…”