Lago x imaging system

BLI was performed by using a Lago X imaging system (Spectral Instruments Imaging). Mice infected with luciferase-expressing XEN36 S. aureus were anesthetized with 2.5% isoflurane, and bioluminescent signals were quantified by using Aura software (Spectral Instruments Imaging). The acquisition parameters were adjusted as follows; exposure time 30 seconds, binning low (2 (link)), f/stop 1.2, and FOV 25. The luminescence data was analyzed using Aura software.

We created the mouse model as mentioned above. After 2 h, mice were intravenously injected with mAb-TPCA-1@HCNPs (n = 4), and the dose of DiR was 2 mg/kg. In addition, healthy mice and model mice intravenously injected with free DiR and isotype IgG-DiR@HCNPs were used as control groups. After 24 h, mice were euthanized, and their organs were collected. Subsequently, the fluorescence intensity of the organs was detected on a Lago X Imaging System (Spectral Instruments Imaging, USA). For the histological study, model mice were intravenously injected with mAb-C6@HCNPs to label NPs. Then, we collected the lungs, fixed them in 4% paraformaldehyde, and embedded them in paraffin. After a series of workflows, we used a CD68 primary antibody followed by cyanine 5-conjugated secondary antibody incubation to detect macrophages. All slides were analyzed using a fluorescence microscope (Olmypus, Tokyo, Japan).

We substituted DiR for TPCA-1 to evaluate the pharmacokinetic profile of mAb-TPCA-1@HCNPs after intravenous (i.v.) injection in mice, and the dose of DiR was 2 mg/kg. At predetermined time points, an equivalent volume of peripheral blood samples was collected from the retro-orbital vein and placed in ninety-six-well plates. Subsequently, the mean fluorescence intensity (MFI) of the samples was determined using a Lago X Imaging System (Spectral Instruments Imaging, Tucson, AZ, USA).