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Visualizing three-dimensional fungal growth using light sheet fluorescence microscopy.

Braulio Gutiérrez-Medina1, Alexis Vázquez-Villa1

  • 1Division of Advanced Materials, Instituto Potosino de Investigación Científica y Tecnológica, Camino a la Presa San José 2055, 78216 San Luis Potosí, Mexico.

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Summary

Researchers developed a novel light-sheet fluorescence microscope (LSFM) for 4D imaging, enabling detailed observation of fungal growth dynamics in three dimensions. This tool visualizes mycelial architecture and growth rates at single-hypha resolution.

Keywords:
3D microscopyHyphal growthLight-sheet fluorescence microscopyMycelial architectureMycelial dynamics

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Area of Science:

  • Mycology
  • Microscopy
  • Biophysics

Background:

  • Fungal morphology is key to understanding growth and environmental interactions.
  • Previous studies of fungal growth were limited to 2D due to a lack of advanced imaging tools for complex 3D structures.

Purpose of the Study:

  • To construct and utilize a light-sheet fluorescence microscope (LSFM) for time-lapse 4D microscopy of fungal growth.
  • To overcome limitations in visualizing and analyzing complex 3D fungal structures.

Main Methods:

  • Construction of a two-color light-sheet fluorescence microscope (LSFM) with detailed optical, electronic, and computational procedures.
  • Time-lapse imaging of fungal strains (Trichoderma atroviride, Neurospora crassa) in liquid media over ~12 hours.
  • Utilizing both autofluorescence and specific tagging for imaging.

Main Results:

  • Successful 4D imaging of fungal growth dynamics at single-hypha resolution within volumes up to ~400 × 1500 × 800 μm³.
  • Visualization of mycelial architecture, hyphal interactions, and measurement of 3D apical extension rates.
  • Demonstrated the capability of LSFM for detailed morphological analysis of fungi.

Conclusions:

  • LSFM provides a powerful new tool for 3D morphological analysis of fungi, from individual hyphae to entire mycelia.
  • This advanced imaging technique facilitates the study of fungal dynamics in complex 3D environments.