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Development of an imaging flow cytometry method for fungal cytological profiling and its potential application in antifungal drug development

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McMahon, Courtney L., Esqueda, Marisol, Yu, Jieh-Juen, Wall, Gina, Romo, Jesus A., Vila, Taissa, Chaturvedi, Ashok, Lopez-Ribot, Jose L. +2 more·2023·Journal of Fungi
DOI 10.3390/jof9070722PMC PMC10381375
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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® 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.

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Authors
McMahon, C. L., Esqueda, M., Yu, J.-J., Wall, G., Romo, J. A., Vila, T., Chaturvedi, A., Lopez-Ribot, J. L., Wormley, F., & Hung, C.-Y.
Affiliations
  • The University of Texas at San Antonio (US)
  • Department of Molecular Microbiology and Immunology and South Texas Center for Emerging Infectious Diseases, The University of Texas at San Antonio, San Antonio, TX 78249, USA
Corresponding author
Hung, Chiung-Yu
ORCID
Venue
Journal of Fungi
Publisher
MDPI AG
Published
2023-06-30
Volume
9
Issue
7
Pages
722
ISSN
2309-608X (ISSN-L)
DOI
10.3390/jof9070722
PMC
PMC10381375
OpenAlex
W4382940804
Cited by
13
References
4
Citations by year
2026: 4 · 2025: 5 · 2024: 4
FWCI
1.57
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Fungi (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • NIH (R21 AI149015, R01 AI135005, U19AI166761, R33AI140823)
  • Margaret Batts Tobin Foundation, San Antonio, TX (R21 AI149015, R01 AI135005, U19AI166761, R33AI140823)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
  • Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Keywords
imaging flow cytometry, antifungal drug development, antifungal screening, Candida albicans, Cryptococcus neoformans, calcofluor white, fluconazole, amphotericin B, caspofungin, fungal cell wall, fungal membranes, phenotypic profiling

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