Ploidy, the number of sets of homologous chromosomes in a cell, can alter cellular physiology, gene regulation, and the spectrum of acquired mutations. Advances in single-cell flow cytometry have greatly improved the understanding of how genome size contributes to diverse biological processes including speciation, adaptation, pathogenesis, and tumorigenesis. For example, fungal pathogens can undergo whole genome duplications during infection of the human host and during acquisition of antifungal drug resistance. Quantification of ploidy is dramatically affected by the nucleic acid staining technique and the flow cytometry analysis of single cells. Ploidy in fungi is also impacted by samples that are heterogeneous for both ploidy and morphology, and control strains with known ploidy must be included in every flow cytometry experiment. To detect ploidy changes within fungal strains, the following protocol was developed to accurately and dependably interrogate single-cell ploidy.
Bibliographical noteFunding Information:
The authors are grateful to Dr. Greg Perry and the Flow Cytometry Core Facility at Creighton University, and support from NIH (3R01GM102487-03S1) for the Yeti flow cytometer. The authors are thankful for various protocols obtained from colleagues including Dr. Aleeza Gerstein and Dr. Judith Berman. Support for this research was provided by LB692 Nebraska Tobacco Settlement Biomedical Research Development New Initiative Grant and Nebraska Established Pro-
- flow cytometry
- genome size