Determination of ploidy in Cryptococcus neoformans by flow cytometry.
We determined the ploidy of Cryptococcus neoformans (28 strains) isolated from patients and nature. The cellular DNA content of these strains, which stained with propidium iodide in comparison to that of two authentic haploid strains, was determined by flow cytometry. All the strains exhibited diphasic histograms. In case of the authentic haploid strains, the first peak was centred around channel 9, and the second peak around channel 18. Most strains exhibited this type of histogram. Some strains exhibited another type of histogram: the first peak was centred around channel 18, and the second one around channel 35. In flow cytometry, the channel number is correlated with the intensity of fluorescence, namely, in proportion to the channel number the DNA content in the cells increases. The cellular DNA content of the second type of histograms showed twice that of the authentic haploid strains, and thus, five of 28 isolates were concluded to be diploid, and the others haploid.
Department of Fungal Infections, Chiba University, Japan.
R
R
Tanaka
H
H
Taguchi
K
K
Takeo
M
M
Miyaji
K
K
Nishimura
D003453:Cryptococcosis / Q000382:microbiology; D003455:Cryptococcus neoformans / Q000166:cytology / Q000235:genetics* / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis; D005434:Flow Cytometry; D006801:Humans; D011003:Ploidies
D004271:DNA, Fungal