To induce peritonitis, male BALB/c mice (6 to 8 weeks old) were injected intraperitoneally with 0.5 ml of sterile zymosan solution (2 mg/ml). At 5.5 hours after challenge, each mouse was intravenously injected with 0.1 ml of saline containing CLP nanoparticles [50 mg/ml (i.e., 10.8 mM)] or luminol at the same dose of the luminol unit. For CLP nanoparticles, the dose was equivalent to 250 mg/kg. The luminescence images were acquired after 30 min (exposure time = 5 min, f/stop = 1, binning = 8, FOV = 21.8 cm). In separate experiments, after 6 hours of stimulation with zymosan, mice were intraperitoneally injected with 0.1 ml of saline containing CLP nanoparticles (50 mg/ml) or luminol at the same dose of the luminol unit. Immediately, bioluminescence imaging was performed. The quantitative analysis was conducted as aforementioned.

To verify cellular distribution of CLP nanoparticles in the abdominal cavity, mice were euthanized and injected with 5 ml of ice-cold PBS. Cells in the peritoneal exudates were separated, stained with anti-mouse V450-CD11b and PE-Ly-6G antibodies, and resuspended in PBS for detection by flow cytometry.

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