Plan apochromat 63x 1.4 ph3 oil objective

Confocal acquisition was performed on a Zeiss Axiovert 200M inverted microscope equipped with a Yokogawa CSU-22 confocal head and a Hamamatsu C9100-02 EM-CCD camera. Images were taken with a Zeiss Plan Apochromat 63x/1.4 Ph3 Oil objective. Improvision Volocity software was used for image acquisition, quantification and co-localization analysis.
For quantifying the distribution of the labelled components in the biofilm, 5 random positions were recorded per condition with a z-spacing of 0.5 μm. The fluorescent structure was detected in every plane and the occupied area determined. Stacks were aligned and the obtained values plotted against the distance from the coverslip. Experiments were performed 2 times.

S. aureus isolates were grown overnight in 300 μl TSB under static conditions in six-well cell culture plates (μ-Dish, Ibidi, Munich, Germany). In some experiments Alexa-568 labelled Wheat germ agglutinin or antibodies against PIA were added to the growth medium. Non-adherent cells were removed by washing with PBS and bacteria were stained using live staining (Live/dead staining, Molecular Probes). Confocal image acquisition was performed on a Zeiss Axiovert 200M inverted microscope equipped with a Yokogawa CSU-22 confocal head and a Hamamatsu C9100-02 EM-CCD camera. Images were taken with a Zeiss Plan Apochromat 63x/1.4 Ph3 Oil objective. Improvision Velocity software was used for image acquisition and quantification.