Microfabrication and its use in investigating fungal biology

Abstract

For nearly 400 years, microscopes have enabled us to explore and describe the diversity of fungi and to observe the effect of experimental manipulations of the cell population. In the last few decades, three new technologies have transformed our application of microscopes in understanding fungal biology and real-time processes in individual cells. First, computer programmable light microscopes enabled real-time imaging of living cells over periods of hours and days to follow temporal processes; second, molecular genetics enabled us to alter gene expression and track fluorescently tagged proteins and reporters inside cells to understand mechanisms. Third, transferred from the semi-conductor industry, microfabrication methods now allow us to manipulate the physical and chemical environment of individual living cells in real-time in situ on the microscope. This "labon-a-chip" technology is now extending into complex multi-cell and compartmentalized environments in "organ-on-a-chip" applications (Last et al., 2021). The contribution of microfabricated surfaces to our understanding of the biology and interactions of fungal cells is just beginning. This review will discuss the contribution that microfabrication has made to the study of biologic processes of yeast, the exploratory growth of hyphae, and how the viscoelastic properties of soft polymer "chips" have been exploited to quantify fungal force and its impact on fungal morphology.

A microfluidic device for simultaneous detection of enzyme secretion and elongation of a single hypha

“…Microfluidics is an excellent tool for manipulating and analyzing single cells as it can confine cells within microstructures (Zare and Kim, 2010;Lecault et al, 2012;Bedekovic and Brand, 2021). Several microfluidic devices have been reported for observing single mycelia.…”

Emerging technologies in microbiology

“…Single cell analyses have become important for understanding the transition of many types of microbes from benign to virulent, free-living to community and population heterogeneity in general. In fungal biology, this has been greatly advanced by microfluidic and microfabrication technologies, that when coupled with molecular genetics and fluorescence imaging capabilities allow the observation of the life cycle of a single yeast cell or filamentous fungi in complex environments (Bedekovic & Brand, 2022).…”

Automated high-content image-based characterization of microorganism behavioral diversity and distribution

“…Drivers acting on time scales of seconds to minutes also play key roles in microbiota assembly and their characterization is required. For example, they may determine heterogeneity in spatial dispersions of species, as in latent stage versus spore germination, or in the displacement and guidance of swimming cells [8] , [9] , [10] . Within this time frame, microorganisms can sense and respond (or not) to drivers and change their distribution rapidly, driven by a variety of physical and (bio)chemical stimuli, such as light, electric fields, and nutrient availability [8] .…”