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The colony forming units (CFU) counting assay is a widely used technique for estimating the number of viable fungal cells in any sample. This method takes advantage of the unique ability of individual fungal cells to proliferate and form visible colonies on agar plates, providing a reliable measure of their reproductive capacity (Meyer et al, Nat Microbiol 8:2304–2314, 2023). The procedure begins with cultivating a fungal culture in a suitable liquid medium. Serial dilutions of the sample are then performed to ensure that the number of colonies formed on the agar plates remains within the optimal range of 30–300 per plate, which is essential for accurate counting. Once the dilutions are prepared, the diluted samples are plated onto specially formulated agar media and incubated at a temperature conducive to fungal growth. During the incubation period, the fungal cells multiply and form distinct colonies. After a predetermined incubation period, the colonies are counted and the concentration of viable fungal cells is expressed as CFU/mL, which is calculated based on the dilution factor (DF) and the volume of the plated sample. The CFU assay is critical in many applications, including fungal susceptibility testing, where it helps to determine the efficacy of antifungal drugs against specific fungal strains (Sahu SR et al, Bio Protoc 13:e4872, 2023). It is also central to pathogenicity assessment and helps researchers understand the virulence of fungal pathogens. In addition, the CFU assay is crucial for quantifying the microbial load of environmental samples, contributing to studies on biodiversity and ecosystem health. Its simplicity, accuracy and adaptability to different fungal species make it an indispensable tool in fungal microbiology, enabling advances in research and clinical diagnostics. Overall, the CFU counting assay remains a cornerstone of mycology, enabling researchers to elucidate complex interactions within fungal communities and their environments (Sieuwerts et al, Lett Appl Microbiol 47:275–278, 2008).

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Counting Colony Forming Units (CFU) to Estimate Viable Fungal Cells in a Sample

  • 章节

  • 发表日期：

    2025年

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  • Ritu Pasrija,
  • Deepika Kumari

摘要

The colony forming units (CFU) counting assay is a widely used technique for estimating the number of viable fungal cells in any sample. This method takes advantage of the unique ability of individual fungal cells to proliferate and form visible colonies on agar plates, providing a reliable measure of their reproductive capacity (Meyer et al, Nat Microbiol 8:2304–2314, 2023). The procedure begins with cultivating a fungal culture in a suitable liquid medium. Serial dilutions of the sample are then performed to ensure that the number of colonies formed on the agar plates remains within the optimal range of 30–300 per plate, which is essential for accurate counting. Once the dilutions are prepared, the diluted samples are plated onto specially formulated agar media and incubated at a temperature conducive to fungal growth. During the incubation period, the fungal cells multiply and form distinct colonies. After a predetermined incubation period, the colonies are counted and the concentration of viable fungal cells is expressed as CFU/mL, which is calculated based on the dilution factor (DF) and the volume of the plated sample. The CFU assay is critical in many applications, including fungal susceptibility testing, where it helps to determine the efficacy of antifungal drugs against specific fungal strains (Sahu SR et al, Bio Protoc 13:e4872, 2023). It is also central to pathogenicity assessment and helps researchers understand the virulence of fungal pathogens. In addition, the CFU assay is crucial for quantifying the microbial load of environmental samples, contributing to studies on biodiversity and ecosystem health. Its simplicity, accuracy and adaptability to different fungal species make it an indispensable tool in fungal microbiology, enabling advances in research and clinical diagnostics. Overall, the CFU counting assay remains a cornerstone of mycology, enabling researchers to elucidate complex interactions within fungal communities and their environments (Sieuwerts et al, Lett Appl Microbiol 47:275–278, 2008).