Abstract
This paper reports the fabrication and application of a Lab-on-a-Chip platform containing single-elastomeric micropillars in channel constrictions, which enable the measurement of protrusive forces exerted by individual fungal hyphae. We show the device design, the fabrication process, and photoresist optimization required to adapt the microfluidic platform to relatively thin hyphae. To demonstrate the applicability of the devices, the oomycete Achlya bisexualis and the fungus Neurospora crassa were cultured on PDMS chips. Devices were combined with confocal imaging to study the interaction of A. bisexualis hyphae with the measurement pillars. The force exerted by individual hyphae of N. crassa was measured and compared with a hyphal growth rate and diameter. The platform provides a new tool to help understand the molecular processes that underlie protrusive growth and this may present new ways to tackle the pathogenic growth of these organisms and thus combat the loss of diversity that they cause. This paper is based on the conference proceedings presented at the 31st IEEE International Conference on Micro Electro Mechanical Systems (MEMS 2018), Belfast. [2018-0090]
Highlights
F UNGI and oomycete are crucial components of most ecosystems, playing a key role in the breakdown of organic matter and nutrient recycling
Based on devices that were originally designed to study nematode movements [16], [17], we have recently reported the use of elastomeric micropillars as a platform technology for the study of protrusive forces generated in hyphae [1], [18]–[20]
Elastomeric micropillars represent a versatile platform technology for the study of protrusive forces generated by hyphal organisms
Summary
F UNGI and oomycete are crucial components of most ecosystems, playing a key role in the breakdown of organic matter and nutrient recycling. This paper has supplementary downloadable material available at http://ieeexplore.ieee.org provided by the authors. In many cases pathogenic growth involves the invasive penetration of host tissue by individual hyphae which extend by the process of tip growth [6]. Previous measurements of protrusive forces exerted by single hyphae have been made using a miniature silicon bridge strain gauge [11]. The current work extends this by optimizing the fabrication and photolithography in order to decrease the sensor pillar diameter and channel-pillar gap from dimensions used with the oomycete Achlya bisexualis, to make the platform suitable for use with fungi with thinner hyphae, such as the model organism N. crassa [21]. Interaction of A. bisexualis hyphae with measurement pillars using confocal microscopy and recorded force measurements of multiple hyphae of N. crassa
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