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A comparative study of four different staining methods for estimation of live yeast form cells ofParacoccidioides brasiliensis

  • Human And Animal Mycology
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Abstract

A comparative study of four different staining methods for estimation of live yeast form cells ofParacoccidioides brasiliensis was carried out. The staining methods used were fluorescent staining, vital dye exclusion tests with erythrosin B and by Janus green and lactophenol cotton blue staining. Colony forming units (cfu) of the yeast form of eightP. brasiliensis isolates on brain heart infusion agar (BHIA) supplemented with 4% horse serum plus 5%P. brasiliensis cell extract (BHIA + HS + EXT) were examined for reliability of staining in determining the number of live fungal units in eight different isolates. Cfu on BHIA + HS + EXT plates showed an excellent plating efficiency over 96% in all isolates tested. The percentage of the live cells indicated by fluorescent staining (FL) or vital dye exclusion test with erythrosin B (EB) or Janus green (JG-1) was lower than that of cfu. By contrast, the percentage due to modified dye exclusion test with Janus green (JG-2) and that due to lactophenol cotton blue staining (LPCB) showed a close correration to that of cfu. Our results indicate that the modified dye exclusion test with Janus green and lactophenol cotton blue staining are useful for estimating cell viability of yeast form cells ofP. brasiliensis.

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Author notes

  1. R. Coelho

    Present address: Department of Pathology, Faculty of Medicine, UNESP, 18618-000, Botucatu, São Paulo, Brazil

Authors and Affiliations

  1. Department of Fungal Infections, Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, 1-8-1 Inohana, Chuo-ku, 260, Chiba, Japan

    Ayako Sano, Nobuyuki Kurita, Kunie Iabuki, R. Coelho, Kanji Takeo, Kazuko Nishimura & Makoto Miyaji

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  1. Ayako Sano
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  2. Nobuyuki Kurita
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  3. Kunie Iabuki
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  4. R. Coelho
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  5. Kanji Takeo
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  6. Kazuko Nishimura
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  7. Makoto Miyaji
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Sano, A., Kurita, N., Iabuki, K. et al. A comparative study of four different staining methods for estimation of live yeast form cells ofParacoccidioides brasiliensis . Mycopathologia 124, 157–161 (1993). https://doi.org/10.1007/BF01103733

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  • DOI: https://doi.org/10.1007/BF01103733

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