Flow cytometric, phase-resolved fluorescence measurement of propidium iodide uptake in macrophages containing phagocytized fluorescent microspheres

ABSTRACT LY303366 is a semisynthetic analog of the antifungal lipopeptide echinocandin B that inhibits (1,3)-β-d-glucan synthase and exhibits efficacy in animal models of human fungal infections. In this study, we utilized flow cytometric analysis of propidium iodide uptake, single-cell sorting, and standard microbiological plating methods to study the antifungal effect of LY303366 on Saccharomyces cerevisiae and Candida albicans. Our data indicate that an initial 5-min pulse treatment with LY303366 caused yeasts to take up propidium iodide and lose their ability to grow. Amphotericin B and cilofungin required longer exposure periods (30 and 180 min, respectively) and higher concentrations to elicit these fungicidal effects. These two measurements of fungicidal activity by LY303366 were highly correlated (r > 0.99) in concentration response and time course experiments. As further validation, LY303366-treated yeasts that stained with propidium iodide were unable to grow in single-cell-sorted cultures. Our data indicate that LY303366 is potent and rapidly fungicidal for actively growing yeasts. The potency and rapid action of this new fungicidal compound suggest that LY303366 may be useful for antifungal therapy.

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Flow cytometric evaluation of DNA stainability with propidium iodide after histone H1 extraction

Cytometry ◽

10.1002/cyto.990100609 ◽

1989 ◽

Vol 10 (6) ◽

pp. 726-730 ◽

Cited By ~ 14

Author(s):

Maria Chiara Giangarè ◽

Ennio Prosperi ◽

Guido Pedrali-Noy ◽

Giovanni Bottiroli

Keyword(s):

Propidium Iodide ◽

Histone H1 ◽

Flow Cytometric ◽

Flow Cytometric Evaluation

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Chromosome doubling in Cattleya tigrina A. Rich

Scientia Plena ◽

10.14808/sci.plena.2019.110202 ◽

2019 ◽

Vol 15 (11) ◽

Author(s):

Thays Saynara Alves Menezes-Sá ◽

Maria de Fátima Arrigoni-Blank ◽

Andréa Santos da Costa ◽

Janay De Almeida Santos-Serejo ◽

Arie Fitzgerald Blank ◽

...

Keyword(s):

Flow Cytometry ◽

Propidium Iodide ◽

Chromosome Doubling ◽

Flow Cytometric Analysis ◽

Leaf Tissue ◽

Stomatal Index ◽

Chromosome Duplication ◽

Flow Cytometric ◽

Young Leaves ◽

Commercial Value

Chromosome doubling induction in orchids may benefit their production for resulting in flowers of higher commercial value, larger size and higher content of substances that intensify the color and fragrance when compared with diploid orchids. This work aimed to induce and confirm artificial polyploidization, using flow cytometry and stomatal analysis. Explants were treated with colchicine at concentrations of 0, 2.5, 7.5, and 12.5 mM, for 24 and 48 hours and with oryzalin, at concentrations of 0, 10, 30, and 50 μM, for three and six days. For the flow cytometric analysis, a sample of leaf tissue was removed from each plant, crushed to release the nuclei and stained with propidium iodide. In addition to flow cytometry, the ploidy of the antimitotic treated plants was evaluated by stomata analysis. Young leaves were used where the density, functionality and stomatal index were evaluated. Colchicine provided induction of satisfactory polyploidy in C. tigrina at all concentrations and times of exposure, obtaining a greater number of polyploid individuals in the concentration of 12.5 mM for 48 hours. Oryzalin did not induce chromosome duplication at the tested concentrations.

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