The lowering of encapsulation was best if the bacteria were in personal direct contact with the host cell membrane. The inset in Fig. 7G shows an ultra-thin section of an adherent A66 cell embedded by using the LRR fixation protocol, which confirmed the loss of the c-Met Inhibitor capsular substance. Capsule structures were shown by all other pneumococci in this chain, as did a pneumococcal chain grown for 2 h in DMEM. The quantity of polysaccharide capsule was significantly reduced for all adherent pneumococci of in connection with the host cell, as shown in Fig. 7L. Three hours after infection just the pneumococci in close contact with the host cell membrane were devoid of capsular structure, while the pneumococci in the connected chain expressed a thick layer of capsule. Pneumococci grown in DMEM again confirmed expression of capsular material over the entire sequence. Ultra-thin section evaluation again indicated that adherent bacteria lost the capsular structure, although bacteria not in close contact still kept the capsule. When infected host cells were treated Eumycetoma with 0. 05% Triton X 100 for 5 min followed by LRR fixation, we could see connected and penetrating pneumococcal stores. As deduced from Fig. 7N, adherent pneumococci in close contact with the host cell membrane and invading pneumococci did not show a visible capsular structure, while pneumococci not in close contact with the host membrane showed the conventional capsular structure after LRR fixation and preparation for FESEM. Pneumococci residing inside host cells showed no detectable capsular polysaccharide material. This means that pneumococci that are in close contact with the cells and show a decreased quantity of capsular polysaccharide aren’t representatives of acapsular mutants that might have been enriched during infection and development. Unpleasant bacteria recovered from epithelial cells by the gentamicin analysis may nevertheless represent natural mutants enriched during growth. Mice were intranasally challenged with S. pneumoniae the lungs, and serotype 3 tension A66 ubiquitin conjugation were processed for morphological analysis from the LRR fixation method and set in a acrylic resin. As shown in Fig. 8, pneumococci were localized in spatial distance from the cells and in touch with lung epithelial tissue. The LRR fixation effectively stabilized and preserved the polysaccharide capsule of pneumococci in lung tissue. FESEM mentioned that pneumococci expressed the tablet in the environment of the lung tissue, although bacteria which were touching lung epithelial tissue showed a severe decrease in the density of the capsular polysaccharide layer. These in vivo results obtained with a mouse model give further evidence for the statement that the total amount of polysaccharide of pneumococci in close contact with host cells is reduced.