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Fig. 2 | BMC Research Notes

Fig. 2

From: Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae

Fig. 2

Indirect immunofluorescent staining of Vero cells infected with R. slovaca and comparison of four fixative solutions. We employed an anti-R. slovaca rabbit serum as primary antibody and goat anti-rabbit Rhodamine Red-X secondary antibody (red fluorescence signal) to probe bacteria. To visualize “comet-tail” formation (in white circles), we applied Alexa Fluor 488 phalloidin for F-actin staining (green fluorescence signal). Rickettsia slovaca formed actin-tails at one pole of the bacterium around 5 µm in length. Likewise, to infected Vero cells with a related intracellular bacterium R. akari, we paralleled fixation step with 3.7% formaldehyde, 4% paraformaldehyde, 4% paraformaldehyde in the cytoskeletal buffer and 4% paraformaldehyde in PHEM buffer. We counterstained nuclei with mounting medium containing DAPI (blue fluorescence signal). Scale bar represents 10 µm

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