Injection into and extraction from single fungal cells

Guillaume-Gentil et al. describe a method that employs a modified AFM tip for selectively sampling from and injecting into individual fungal cells of differing morphology. The authors describe extensive modifications on their system previously used for mammalian cells to overcome many of the challenges associated with working on single fungal cells. The direct delivery of molecules and the sampling of endogenous compounds into and from living cells provide powerful means to modulate and study cellular functions. Intracellular injection and extraction remain challenging for fungal cells that possess a cell wall. The most common methods for intracellular delivery into fungi rely on the initial degradation of the cell wall to generate protoplasts, a step that represents a major bottleneck in terms of time, efficiency, standardization, and cell viability. Here, we show that fluidic force microscopy enables the injection of solutions and cytoplasmic fluid extraction into and out of individual fungal cells, including unicellular model yeasts and multicellular filamentous fungi. The approach is strain- and cargo-independent and opens new opportunities for manipulating and analyzing fungi. We also perturb individual hyphal compartments within intact mycelial networks to study the cellular response at the single cell level.

Automated summary

Guillaume-Gentil et al. present a method that utilizes a modified AFM tip to selectively sample and inject into fungal cells of different morphology. The authors made extensive modifications on their existing system for mammalian cells to overcome challenges encountered when working on single fungal cells. They demonstrate the effectiveness of fluidic force microscopy in injecting solutions and extracting cytoplasmic fluid from individual fungal cells, providing new opportunities for manipulating and analyzing fungi.