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. 2011 Oct;79(10):3872–3886. doi: 10.1128/IAI.05631-11

Fig. 6.

Fig. 6.

Fluorescence in situ hybridization. FISH was performed using a trypsinized culture of HeLa cells (5 × 103 cells/ml) grown on coverslips. Probe-hybridized slides were air dried, mounted using the SlowFade Gold antifade reagent, and visualized using an inverted Axio Observer Z1 confocal microscope. The images were analyzed using Zeiss LSM 710 software. The images shown are representative of more than 12 viewing fields under a given magnification. F. alocis D-62D and P. gingivalis W83 were used in this study. (A) P. gingivalis W83 control cells showing specific red-orange fluorescence (645-nm absorbance/665-nm emission) (magnification, ×20). (B) F. alocis D-62D control cells showing specific green fluorescence (549-nm absorbance/563-nm emission) (magnification, ×20). (C) Negative control showing no fluorescence using DAPI (magnifi- cation, ×20). (D) Coinfection producing marginal green fluorescence around the epithelial cells, showing adherence of F. alocis D-62D seen after 20 min p.i. (magnification, ×40). (E) Coinfection producing a shift in the fluorescence, showing invasion by P. gingivalis W83 over F. alocis D-62D at 30 min p.i. (magnification, ×40). (F) Coinfected epithelial cells showing peripheral adherence of F. alocis D-62D and early invasion by P. gingivalis W83 (magnification, ×40) at 45 min p.i. (G) Coinfected individual epithelial cell showing early invasion by P. gingivalis W83 (magnification, ×63) at 45 min p.i. (H) Coinfected individual epithelial cell showing early invasion by P. gingivalis W83 (intense red-orange fluorescence in the cytoplasm) (magnification, ×40) at 45 min p.i. (I) Coinfected epithelial cell showing sparse adherence of F. alocis D-62D and cytoplasm showing the invading P. gingivalis W83 (magnification, ×40) at 45 min p.i. (J) Coinfection of F. alocis D-62D and P. gingivalis W83 showing generalized invasion of epithelial cells at 1 h p.i. (magnification, ×20). (K) Coinfection of F. alocis D-62D and P. gingivalis W83 showing general invasion of epithelial cells at 1 h p.i. (magnification, ×40). (L and M) Equal invasion was noticed after 1 h p.i. (magnification, ×20). (N) Invasion by P. gingivalis W83 monoculture in epithelial cells (magnification, ×20). (O) Invasion by F. alocis monoculture in epithelial cells (magnification, ×20).