A Rapid Colorimetric Assay for Determination of Leukocyte-Mediated Inhibition of Mycelial Growth of Candida albicans

• Published in: MICROBIOLOGY and IMMUNOLOGY Vol. 38; no. 5; pp. 385 - 388
• Main Authors: Abe, Shigeru, Satoh, Tamae, Tokuda, Yoshiko, Tansho, Shigeru, Yamaguchi, Hideyo
• Format: Journal Article
• Language: English
• Published: Tokyo Blackwell Publishing Ltd 01.01.1994; Center For Academic Publications Japan; Center for Academic Publications Japan
• Subjects: Animals; Biological and medical sciences; Candida albicans; Candida albicans - growth & development; Candida albicans - immunology; Colorimetry - methods; Crystal violet; Fundamental and applied biological sciences. Psychology; Gentian Violet; Glucose - metabolism; Growth inhibition assay; Leukocytes; Macrophages - immunology; Mice; Mice, Inbred C3H; Microbiology; Mycological methods and techniques used in mycology; Mycology; Neutrophils - immunology; Fungi; Candida albicans; Microorganism growth; Yeast; Investigation method; Leukocyte; Colorimetry; Fungi Imperfecti; Inhibition; Thallophyta
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1994.tb01795.x

A staining method with crystal violet (CV) was demonstrated to be useful for a simple, quick and objective assessment of in vitro growth inhibitory activity of leukocytes against Candida albicans cells. Candida cells incubated with murine neutrophils or macrophages for 14hr in microwells were stained with CV and, after washing with 0.25% sodium dodecyl sulfate (SDS), treated with isopropanol containing HCl (0.04N) to extract Candida cell-bound CV. Then the absorbance at 590nm of the isopropanol extract was photometrically measured. The results showed that the photometrical absorbance was proportional to the amount of 3H-glucose taken up by C. albicans cells, which reflected the number of viable Candida cells.