Laboratory of Cellular Biology, Department of Biology, ICB, Federal University of Juiz de Fora, UFJF, Juiz de Fora, MG, Brazil; Faculty of Medical Sciences, Radboud University, Nijmegen, the Netherlands.
Laboratory of Cellular Biology, Department of Biology, ICB, Federal University of Juiz de Fora, UFJF, Juiz de Fora, MG, Brazil.
Microbiol Res. 2021 Sep;250:126806. doi: 10.1016/j.micres.2021.126806. Epub 2021 Jun 10.
New strategies that enable fast and accurate visualization of Candida biofilms are necessary to better study their structure and response to antifungals agents. Here, we applied whole slide imaging (WSI) to study biofilm formation of Candida species. Three relevant biofilm-forming Candida species (C. albicans ATCC 10231, C. glabrata ATCC 2001, and C. tropicalis ATCC 750) were cultivated on glass coverslips both in presence and absence of widely used antifungals. Accumulated biofilms were stained with fluorescent markers and scanned in both bright-field and fluorescence modes using a WSI digital scanner. WSI enabled clear assessment of both size and structural features of Candida biofilms. Quantitative analyses readily detected reductions in biofilm-covered surface area upon antifungal exposure. Furthermore, we show that the overall biofilm growth can be adequately assessed across both bright-field and fluorescence modes. At the single-cell level, WSI proved adequate, as morphometric parameters evaluated with WSI did not differ significantly from those obtained with scanning electron microscopy, considered as golden standard at single-cell resolution. Thus, WSI allows for reliable visualization of Candida biofilms enabling both large-scale growth assessment and morphometric characterization of single-cell features, making it an important addition to the available microscopic toolset to image and analyse fungal biofilm growth.
为了更好地研究其结构和对抗真菌药物的反应，需要新的策略来实现对念珠菌生物膜的快速、准确可视化。在这里，我们应用全玻片成像（WSI）来研究念珠菌属物种的生物膜形成。三种相关的生物膜形成念珠菌（白色念珠菌 ATCC 10231、光滑念珠菌 ATCC 2001 和热带念珠菌 ATCC 750）在存在和不存在广泛使用的抗真菌药物的情况下在玻璃载玻片上培养。用荧光标记物对累积的生物膜进行染色，并使用 WSI 数字扫描仪在明场和荧光模式下进行扫描。WSI 能够清晰评估念珠菌生物膜的大小和结构特征。定量分析很容易检测到抗真菌药物暴露后生物膜覆盖表面积的减少。此外，我们表明可以在明场和荧光模式下充分评估整体生物膜生长。在单细胞水平上，WSI 证明是足够的，因为用 WSI 评估的形态参数与用扫描电子显微镜（被认为是单细胞分辨率的金标准）获得的参数没有显著差异。因此，WSI 允许对念珠菌生物膜进行可靠的可视化，从而能够对大规模生长进行评估，并对单细胞特征进行形态计量学表征，这是对现有的用于成像和分析真菌生物膜生长的显微镜工具集的重要补充。
