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Updated: Aug 28, 2025
Candida albicans Biofilm Chip CaBChip for High-throughput Antifungal Drug Screening
Published on: July 18, 2012
Optotracing for live selective fluorescence-based detection of Candida albicans biofilms
Elina Kärkkäinen1,2, Saga G Jakobsson1,3, Ulrica Edlund1,3
1AIMES-Center for the Advancement of Integrated Medical and Engineering Sciences Karolinska Institutet and KTH Royal Institute of Technology, Stockholm, Sweden.
Abstract:
Candida albicans is the most common fungal pathogen in humans, implicated in hospital-acquired infections, secondary infections in human immunodeficiency virus (HIV) patients, and is a significant contributor to the global antimicrobial resistance (AMR) burden. Early detection of this pathogen is needed to guide preventative strategies and the selection and development of therapeutic treatments. Fungal biofilms are a unique heterogeneous mix of cell types, extracellular carbohydrates and amyloid aggregates. Perhaps due to the dominance of carbohydrates in fungi, to date, few specific methods are available for the detection of fungal biofilms. Here we present a new optotracing-based method for the detection and analysis of yeast and biofilms based on C. albicans SC5314 as a model. Using commercial extracts of cell wall carbohydrates, we showed the capability of the optotracer EbbaBiolight 680 for detecting chitin and β-glucans. The sensitivity of this tracer to these carbohydrates in their native environment within fungal cells enabled the visualization of both yeast and hyphal forms of the microbe. Analysis of optotracer fluorescence by confocal laser scanning microscopy revealed extensive staining of fungi cell walls as well as the presence of intracellular amyloid aggregates within a subpopulation of cells within the biofilm. Further analysis of the photophysical properties of bound tracers by spectroscopy and spectral imaging revealed polymorphisms between amyloid aggregates within yeast and hyphal cells and enabled their differentiation. With exceptional spatial and temporal resolution, this assay adds a new technique that facilitates future understanding of fungal biofilms and their formation, and enables direct, unbiased diagnostics of these medically relevant biofilms, as well as the development of antifungal strategies.
Insights
A new optotracing method detects fungal biofilms by visualizing chitin and beta-glucans. This technique aids in diagnosing medically relevant biofilms and developing antifungal strategies against Candida albicans.
Area of Science:
- Mycology
- Biotechnology
- Medical Diagnostics
Background:
- Candida albicans is a major fungal pathogen causing hospital infections and contributing to antimicrobial resistance.
- Fungal biofilms are complex structures with few specific detection methods available.
- Early detection of C. albicans is crucial for effective treatment and prevention.
Purpose of the Study:
- To develop a novel optotracing method for detecting and analyzing yeast and biofilms of Candida albicans.
- To visualize fungal cell wall components and intracellular structures within biofilms.
- To differentiate between yeast and hyphal forms within biofilms using spectral imaging.
Main Methods:
- Utilized the optotracer EbbaBiolight 680 to detect chitin and beta-glucans in C. albicans.
- Employed confocal laser scanning microscopy to visualize tracer fluorescence in fungal cells and biofilms.
- Applied spectroscopy and spectral imaging to analyze photophysical properties of bound tracers.
Main Results:
- Demonstrated EbbaBiolight 680's capability to detect chitin and beta-glucans in C. albicans.
- Visualized yeast and hyphal forms, along with intracellular amyloid aggregates within biofilms.
- Revealed polymorphisms in amyloid aggregates, enabling differentiation between yeast and hyphal cells.
Conclusions:
- The optotracing method provides high spatial and temporal resolution for fungal biofilm analysis.
- This technique enables direct, unbiased diagnostics of medically relevant biofilms.
- Facilitates understanding of biofilm formation and aids in developing antifungal strategies.
