Development of an Imaging Flow Cytometry Method for Fungal Cytological Profiling and Its Potential Application in Antifungal Drug Development

Date
2023-06-30
Authors
McMahon, Courtney L.
Esqueda, Marisol
Yu, Jieh-Juen
Wall, Gina
Romo, Jesus A.
Vila, Taissa
Chaturvedi, Ashok
Lopez-Ribot, Jose L.
Wormley, Floyd
Hung, Chiung-Yu
Journal Title
Journal ISSN
Volume Title
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Abstract

Automated imaging techniques have been in increasing demand for the more advanced analysis and efficient characterization of cellular phenotypes. The success of the image-based profiling method hinges on assays that can rapidly and simultaneously capture a wide range of phenotypic features. We have developed an automated image acquisition method for fungal cytological profiling (FCP) using an imaging flow cytometer that can objectively measure over 250 features of a single fungal cell. Fungal cells were labeled with calcofluor white and FM4-64FX, which bind to the cell wall and lipophilic membrane, respectively. Images of single cells were analyzed using IDEAS(R) software. We first acquired FCPs of fungal cells treated with fluconazole, amphotericin B, and caspofungin, each with a distinct mode of action, to establish FCP databases of profiles associated with specific antifungal treatment. Once fully established, we investigated the potential application of this technique as a screening methodology to identify compounds with novel antifungal activity against Candida albicans and Cryptococcus neoformans. Altogether, we have developed a rapid, powerful, and novel image-profiling method for the phenotypic characterization of fungal cells, also with potential applications in antifungal drug development.

Description
Keywords
antifungal, drug discovery, cytological profiling, imaging flow cytometry, phenotypic screening
Citation
Journal of Fungi 9 (7): 722 (2023)
Department
Molecular Microbiology and Immunology