Methicillin-resistant Staphylococcus aureus (MRSA) causes both hospital- and community-acquired pneumonia, but translatable models to investigate early pathogenesis and potential therapeutics are limited. Precision-cut lung slices (PCLS) represent a novel tool for dynamic time-course studies of MRSA infection with higher throughput than in vivo mouse models. PCLS retain their complex architecture with all resident cells present, including immune cells, making them an effective model of the host environment. Additionally, single mouse or human tissue samples can produce hundreds of lung slices, thus reducing the number of animals required for experiments, and maximising the high-throughput potential of human tissue. To establish infection, PCLS from agarose-inflated lungs of male 8–10-week-old C57BL/6 mice were inoculated with 105 CFU/mL dsRed-labelled MRSA, incubated for 2 h before transfer to fresh media. Infected and uninfected tissue were monitored over 48 h for total tissue viability (LDH-assay), bacterial load, bacterial localisation (fluorescence microscopy), resident immune cell and inflammatory cytokine response (spectral flow cytometry and BioLegend LEGENDplexTM). Viability was similar in uninfected and infected PCLS. Diffuse MRSA colonisation was observed across all structures in PCLS, and CFU/mL increased >100,000 fold by 48 hpi (n=6, P<0.001). MRSA associated with CD45+ immune cells over time (2 hpi 9±2%; 24 hpi 74±10%, n=4, P<0.05), particularly professional phagocytic cell populations such as alveolar macrophages and dendritic cells. TNF-α, GM-CSF and IL-6 were increased in MRSA-infected PCLS within 24 hpi (n=6, P<0.05), suggesting that mouse PCLS retain intact resident immune cells and inflammatory responses over the course of infection. Our findings support the future application of PCLS as a high throughput platform for the assessment of drugs to prevent bacterial infection. Future studies aim to translate our initial findings to ex vivo human lung tissue, investigate the contributions of specific S. aureus proteins to immune cell cytotoxicity, and extend our model to investigate Mycobacterium tuberculosis infection.