MRSA replicates in KCs. (a) SD-IVM image of liver 8 h after i.v. infection with MRSA (MW2-GFP). Arrow, large cluster of bacteria co-localizing with KCs (purple). Bar, 50 µm. (b) Quantification of SD-IVM images for GFP-clusters >50 µm²/FOV assessed at 30 m and 8 h after i.v. infection with MRSA (MW2-GFP). (a and b) n = 5 per time point. Error bars, SEM; **, P < 0.01, Student’s t test. Data were pooled from two independent experiments. (c) 3D SD-IVM image reconstruction of a staphylococcal cluster (MW2-GFP) co-localizing with KCs (F4/80, purple) 8 h after i.v. infection. Bar, 5 µm (Video 3). (d) Number of MRSA (MW2-GFP) per infected KCs at 30 m or 8 h after i.v. infection. (c and d) n = 100 KCs compiled from eight mice per condition. Error bars, SEM; Student’s t test. (e) Quantification of captured Staphylococci (MW2-GFP) over different time intervals after i.v. infection. n = 3 per time interval. Data were pooled from two independent experiments. (f) Flow cytometric analysis of Syto 60 dye upon culturing MRSA (MW2-GFP) WT or heat-inactivated for different time points. One representative out of three independent experiments is shown. (g) SD-IVM image of dilution of Syto 60 by replicating MRSA within KCs. KCs (F4/80, purple) in mouse liver 8 h after infection with MRSA (MW2-GFP; green) in vitro labeled with Syto 60 (blue). White arrow indicates staphylococci that have lost Syto 60 dye as a result of replication. Yellow arrow indicates MRSA that still have Syto 60. Bar, 5 µm. Shown is one representative image out of five independent experiments.