Fig 1: Localization of S. aureus on the border between stratum spinosum and stratum basale. Overview (a) and detailed image (b) of nasal tissue from an S. aureus carrier by confocal laser scanning microscopy. Scale bar is in micrometers. White brace indicates the region which is magnified in (b). Green fluorescence labeling of S. aureus, blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). We used a CLSM ×63 objective
Fig 2: Localization of S. aureus in the lower part of epidermis by CLSM. Overview and detailed image/zoomed portion of the image showing (a) S. aureus on the border between stratum spinosum and stratum basale, and (b) S. aureus on the border between the stratum basale and dermis. Inset represents a zoomed portion of the image. Scale bar is in micrometers. Green fluorescence labeling of S. aureus, blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). We used a CLSM ×63 objective
Fig 3: Localization of S. aureus in the upper part of epidermis. Inset represents a zoomed portion of the image. Scale bar is in micrometers. We used a confocal laser scanning microscopy (CLSM) ×63 objective. a-b S. aureus is localized on the border between stratum corneum and stratum granulosum. Green fluorescence labeling of S. aureus, blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), and DRAQ5 (BioStatus). c S. aureus in stratum corneum. The same labeling is used as in a-b, but in addition, we used Alexa Fluor 594 Phalloidin (A12381, Molecular Probes™, Thermo Fisher Scientific) staining actin red. d Combined CLSM and light microscopy of nasal tissue section. S. aureus seen as red cocci in pairs, presumably on the border between stratum corneum and stratum granulosum. Primary S. aureus rabbit polyclonal antibody (ab20920, Abcam) and a secondary antibody Alexa Fluor 546 goat anti-rabbit IgG (Molecular Probes™, Thermo Fisher Scientific) were used
Fig 4: Confocal laser scanning microscopy of frozen section of nasal tissue from an S. aureus carrier. Overview (a-b) and detailed/zoomed portion of image (c-d) showing S. aureus in stratum spinosum. Scale bar is in micrometers. Green fluorescence labeling of S. aureus, blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA, red fluorescence labeling of actin. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), Alexa Fluor 594 Phalloidin (Molecular Probes™, Thermo Fisher Scientific) and DRAQ5 (BioStatus). We used a CLSM ×63 objective
Fig 5: Localization of S. aureus in stratum spinosum by CLSM. Inset represents a zoomed portion of the image. Scale bar is in micrometers. We used a CLSM ×63 objective. The sections are oriented so that the outermost epidermis layer is shown in the upper left corner. a-b Green fluorescence labeling of S. aureus, blue fluorescence labeling of keratinocyte cell nuclei and S. aureus DNA. c Same labeling as in a and b, but in addition we used red fluorescence labeling of actin. Primary antibody rabbit polyclonal antibody to S. aureus (ab20920, Abcam), secondary antibody Alexa Fluor 488® goat anti-rabbit IgG (Molecular Probe™, Thermo Fisher Scientific), Alexa Fluor 594 Phalloidin (Molecular Probes™, Thermo Fisher Scientific) and DRAQ5 (BioStatus)
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