Quantitative Analysis of Candida Cell Wall Components by Flow Cytometrywith Triple-Fluorescence Staining

MF Nogueira, F Istel, S Jenull, L A Walker, N Gow, T Lion (Corresponding Author)

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Abstract

Detailed analysis of dynamic fungal cell wall components is crucial to our understanding of fungal systematics and the biology and
physiology of fungal growth. In fungal pathogens this is of particular importance in examining their response to stress. However,
current methodologies do not permit fast and accurate or quantitative analysis of cell wall carbohydrate components. Here, we provide
a novel method permitting simultaneous quantitative analysis of the major cell wall components of Candida species relying on triplestaining
with fluorescent labeling of chitin, β-glucans and mannans. Quantification is based on flow cytometry whereas qualitative
analysis can be performed by direct imaging using fluorescence microscopy. We validated the method by determining the in vitro
responses of different Candida species to the challenge of antifungal treatment with caspofungin. The assay facilitated rapid analysis
of adaptive changes in cell wall composition upon caspofungin-induced damage. The method can be exploited for comprehensive
quantitative analysis of Candida cell wall components, with relevant implications for clinical diagnosis of fungal infections in general.
Original languageEnglish
Article number101
Number of pages9
JournalJournal of Microbiology and Modern Techniques
Volume2
Issue number1
Early online date20 Oct 2017
DOIs
Publication statusPublished - 31 Dec 2017

Keywords

  • Candida spp
  • cell wall
  • triple staining
  • chitin
  • glucans
  • mannans
  • flow cytometry

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