Fig. 1

Indirect immunofluorescent staining of Vero cells infected with R. akari and comparison of four fixative solutions. We applied an anti-R. akari rabbit serum as primary antibody and goat anti-rabbit Rhodamine Red-X secondary antibody (red fluorescence signal) to visualize bacteria. To show the overall shape of the host cells and actin polymerization by R. akari (in a white circle), we employed Alexa Fluor 488 phalloidin for F-actin staining (green fluorescence signal). We compared 3.7% formaldehyde, 4% paraformaldehyde, 4% paraformaldehyde in the cytoskeletal buffer and 4% paraformaldehyde in PHEM buffer as fixatives. For nuclear counterstaining, mounting medium with DAPI was used (blue fluorescence signal). Scale bar represents 10 µm