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Published July 1, 2024 | Published
Journal Article Open

Long-term imaging of three-dimensional hyphal development using the ePetri dish

  • 1. ROR icon California Institute of Technology

Abstract

Imaging three-dimensional microbial development and behavior over extended periods is crucial for advancing microbiological studies. Here, we introduce an upgraded ePetri dish system specifically designed for extended microbial culturing and 3D imaging, addressing the limitations of existing methods. Our approach includes a sealed growth chamber to enable long-term culturing, and a multi-step reconstruction algorithm that integrates 3D deconvolution, image filtering, ridge, and skeleton detection for detailed visualization of the hyphal network. The system effectively monitored the development of Aspergillus brasiliensis hyphae over a seven-day period, demonstrating the growth medium’s stability within the chamber. The system’s 3D imaging capability was validated in a volume of 5.5 mm × 4 mm × 0.5 mm, revealing a radial growth pattern of fungal hyphae. Additionally, we show that the system can identify potential filter failures that are undetectable with 2D imaging. With these capabilities, the upgraded ePetri dish represents a significant advancement in long-term 3D microbial imaging, promising new insights into microbial development and behavior across various microbiological research areas.

Copyright and License

© 2024 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.

Data Availability

The code and data underlying this study are available from Mango Inc.

Funding

Mango Inc.

Conflict of Interest

Patent applications covering the ePetri dish hardware and reconstruction algorithm have been filed by Mango Inc.

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Additional details

Created:
June 25, 2024
Modified:
June 25, 2024