Injection into and extraction from single fungal cells

Area of Science:

• Cell Biology
• Mycology
• Biotechnology

Background:

• Direct molecular delivery and compound sampling are crucial for studying cellular functions.
• Fungal cell walls present significant challenges for intracellular manipulation, often requiring protoplast generation.
• Current methods for fungal intracellular delivery are time-consuming, inefficient, and can impact cell viability.

Purpose of the Study:

• To develop a novel method for direct intracellular delivery and extraction in fungal cells.
• To overcome the limitations associated with protoplast generation in fungi.
• To enable precise manipulation and analysis of individual fungal cells and hyphal compartments.

Main Methods:

• Utilized fluidic force microscopy (FFM) for intracellular applications.
• Applied FFM to inject solutions and extract cytoplasmic fluid from individual fungal cells.
• Tested the method on both unicellular yeasts and multicellular filamentous fungi.

Main Results:

• Demonstrated successful intracellular injection and cytoplasmic fluid extraction using FFM in various fungal species.
• Showcased the strain- and cargo-independent nature of the FFM approach.
• Successfully perturbed individual hyphal compartments within intact mycelial networks.

Conclusions:

• Fluidic force microscopy provides a direct, efficient, and versatile method for fungal cell manipulation.
• This technique bypasses the need for cell wall degradation, preserving cell viability and standardization.
• Opens new avenues for research in fungal cell biology, genetics, and biotechnology.