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Updated: Nov 17, 2025
Application of Membrane and Cell Wall Selective Fluorescent Dyes for Live-Cell Imaging of Filamentous Fungi
Published on: November 28, 2019
Fluorescent toys 'n' tools lighting the way in fungal research
Wouter Van Genechten1,2,3, Patrick Van Dijck1,2, Liesbeth Demuyser1,2
1VIB-KU Leuven Center for Microbiology, Kasteelpark Arenberg 31, 3001 Leuven-heverlee, Belgium.
Abstract:
Although largely overlooked compared to bacterial infections, fungal infections pose a significant threat to the health of humans and other organisms. Many pathogenic fungi, especially Candida species, are extremely versatile and flexible in adapting to various host niches and stressful situations. This leads to high pathogenicity and increasing resistance to existing drugs. Due to the high level of conservation between fungi and mammalian cells, it is hard to find fungus-specific drug targets for novel therapy development. In this respect, it is vital to understand how these fungi function on a molecular, cellular as well as organismal level. Fluorescence imaging allows for detailed analysis of molecular mechanisms, cellular structures and interactions on different levels. In this manuscript, we provide researchers with an elaborate and contemporary overview of fluorescence techniques that can be used to study fungal pathogens. We focus on the available fluorescent labelling techniques and guide our readers through the different relevant applications of fluorescent imaging, from subcellular events to multispecies interactions and diagnostics. As well as cautioning researchers for potential challenges and obstacles, we offer hands-on tips and tricks for efficient experimentation and share our expert-view on future developments and possible improvements.
Insights
Fungal infections are a major health threat. This review details fluorescence imaging techniques to study fungal pathogens, aiding in understanding their molecular mechanisms and developing new therapies.
Area of Science:
- Microbiology
- Biotechnology
- Medical Mycology
Background:
- Fungal infections present a significant, yet often underestimated, global health challenge.
- Pathogenic fungi, particularly Candida species, exhibit remarkable adaptability, leading to increased pathogenicity and drug resistance.
- Identifying fungus-specific drug targets is difficult due to conserved cellular machinery between fungi and mammals.
Purpose of the Study:
- To provide a comprehensive overview of fluorescence imaging techniques for studying fungal pathogens.
- To guide researchers in applying these techniques from subcellular to organismal levels.
- To highlight practical tips and future directions in fungal research using fluorescence.
Main Methods:
- Review of contemporary fluorescence labelling and imaging techniques relevant to fungal research.
- Discussion of applications including subcellular dynamics, cellular structures, and host-pathogen interactions.
- Inclusion of practical advice for experimental design and troubleshooting.
Main Results:
- Fluorescence imaging offers powerful tools for dissecting fungal molecular and cellular processes.
- Techniques span from detailed subcellular analysis to studying complex multispecies interactions.
- The manuscript serves as a practical guide for researchers in the field.
Conclusions:
- Understanding fungal pathogens at multiple biological levels is crucial for combating infections.
- Fluorescence imaging is an indispensable methodology for advancing fungal research and drug discovery.
- Continued development of fluorescence techniques promises enhanced insights into fungal biology and pathogenesis.
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