Microfluidics structures for probing the dynamic behaviour of filamentous fungi

Although filamentous fungi live in physically and chemically complex natural environments that require optimal survival strategies, both at colony and individual cell level, their growth dynamics are usually studied on homogenous media. This study proposes a new research methodology based on the purposeful design, fabrication and operation of microfluidics structures to examine the temporal and spatial responses of filamentous fungi. Two model fungal strains, the wild type of Neurospora crassa - a commonly used model organisms - and the ro-1 mutant strain of this species impaired in hyphal growth and morphology, have been chosen to demonstrate the potential of this new methodology. Time-lapse observations of both species show that filamentous fungi respond rapidly to the physically microstructured environment without any detectable temporal or spatial adjustment period. Despite their genetic differences, and consequently different growth behaviour, both strains present efficient space-search strategies enabling them to solve the microsized networks successfully and in similar periods, thus demonstrating that the space-searching algorithms are robust and mutation-independent. Additionally, the use of the proposed methodology could put in evidence new biological mechanisms responsible for the apical extension of filamentous fungi, beyond the classical theory based on the central role of Spitzenkorper.