Presence of bacteria in mucosal biofilms of mice with oropharyngeal candidiasis.

<p>Panels A and B depict tissue sections stained with an anti-<i>Candida</i> pAb (green) and the nucleic acid stain Syto59 (red). Panel B is a 3.5× zoom image of the marked area in Panel A. Notice the close association of <i>C. albicans</i> and bacterial cells (arrows). Panel C depicts a tissue section stained with an anti-<i>Candida</i> antibody (green), processed for fluorescence in situ hybridization (FISH) with the all bacteria-specific oligonucleotide probe EUB388 (red) and counterstained with the nucleic acid stain Hoechst 33258 (blue). Notice the presence of bacteria (pink) throughout the mucosal biofilms (arrows). Scale bar = 20 µm.</p

β-glucan and extracellular material staining during <i>C. albicans</i> SC5314 in vitro biofilm growth on glass.

<p>Panels depict 3D reconstructions of confocal stacks of images of 24h (A), 48h (B) and 72h (C) biofilms of <i>C. albicans</i> grown on cover slips and stained for β-glucan with BFDiv mAb (red) and ConA-Alexa 488 (green). Notice that regardless of biofilm thickness β-glucan is localized in the growing end of the biofilm (arrows).</p

Cytokeratin presence in <i>C. albicans</i> biofilms formed on the tongue of mice with oropharyngeal candidiasis.

<p>Tissue sections in panels A, B and C were stained with anti-cytokeratin mAb (red) and anti-<i>Candida</i> pAb (green). Host cell nuclei were visualized with TO-PRO-3 (blue). Panel D represents an isotype control (IgG) stain. Arrows indicate areas where fungal cells are surrounded by keratin. Scale bar = 20 µm.</p

Neutrophils form aggregates in tongue biofilms of <i>C.albicans</i>-infected mice.

<p>Panels A–D depict tissue sections stained with an anti-mouse neutrophil mAb (red), an anti-<i>Candida</i> Ab (green) and the nucleic acid stain TO-PRO-3 (blue). Panel E depicts a confocal image of a negative control stain (primary anti-neutrophil mAb was omitted). Arrows indicate the presence of neutrophils directly juxtaposed to, or within the biofilm mass. Scale bar = 20 µm.</p

β-glucan and extracellular material staining in <i>C. albicans</i> biofilms forming on glass.

<p>Panel A depicts a 2h biofilm and panel B depicts a 48h biofilm. Biofilms of the GFP-expressing <i>C. albicans</i> strain (green) were stained for β-glucan with a BFDiv monoclonal antibody (red) and the extracellular material was stained with ConA-Alexa 350 (blue). In 2h biofilms there is partial co-localization of the BFDiv mAb and ConA (pink). BFDiv stains parts of the fungal cell, but not the germinating buds, and ConA stains the entire fungal cell surface (3A). In 48h biofilms deposits of cell-dissociated ECM stained with ConA but not with BFDiv (3B, arrows). Scale bar = 20 µm.</p

<i>C. albicans</i> presence in “white plaque” lesions formed on the tongue of mice with oropharyngeal candidiasis.

<p><i>C. albicans</i>-challenged mice were sacrificed after 5 days of oral exposure to the GFP-expressing strain MRL51. Panel A depicts the dorsal aspect of a tongue from an uninfected control. Panel B depicts the white plaque lesions formed on the tongue of an infected mouse. Panel C depicts a three dimensional reconstruction of a live biofilm as visualized via confocal microscopy.</p