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Fluidic force microscopy allows direct molecular delivery and compound sampling in fungal cells without cell wall removal. This innovative technique overcomes previous limitations, enabling precise manipulation and analysis of yeasts and filamentous fungi.

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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.