Automated, continuous video microscopy tracking of hyphal growth

Area of Science:

• Mycology
• Microscopy
• Biophysics

Background:

• Fungal growth, involving hyphal elongation and branching, is crucial but traditionally requires laborious manual image analysis.
• Automated methods are needed to overcome the limitations of manual observation for studying fungal dynamics.

Purpose of the Study:

• To develop and validate a novel, automated system for real-time tracking and quantitative analysis of fungal hyphae growth.
• To investigate the dynamics of hyphal elongation, branching, and morphology in *Trichoderma atroviride*.

Main Methods:

• Computer-controlled microscopy stage with stepper motors for precise sample positioning.
• Real-time image-processing algorithm to detect and track hyphal tips during elongation.
• Automated autofocus correction and field-of-view maintenance for continuous observation.

Main Results:

• Automated, continuous tracking of *Trichoderma atroviride* hyphae over extended periods (up to 14 hours).
• Determination of characteristic hyphal elongation rates (approximately 70 nm/s) and persistence length (362 µm).
• Observation of transient growth arrest events in parental hyphae preceding apical branching.

Conclusions:

• Integration of image analysis and computer control provides a powerful tool for quantitative microscopy of fungal hyphae.
• The automated system enables novel discoveries regarding fungal growth dynamics, including pre-branching arrest events.
• This technology facilitates high-throughput, detailed analysis of fungal morphogenesis and growth kinetics.