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The established dogma termed the <jats:italic>Race for the Surface<\/jats:italic> posits that contaminating bacteria compete with host cells for the implant post\u2010op, which remains unproven without real\u2010time in vivo evidence. Thus, we modified the murine longitudinal intravital imaging of the bone marrow (LIMB) system to allow real\u2010time quantification of green fluorescent protein\u00a0(GFP+) host cells and enhanced cyan fluorescent protein\u00a0(ECFP+) or red fluorescent protein\u00a0(RFP+) methicillin\u2010resistant <jats:italic>Staphylococcus aureus<\/jats:italic> (MRSA) proximal to a transfemoral implant. Following inoculation with ~10<jats:sup>5<\/jats:sup> CFU, an L\u2010shaped metal implant was press\u2010fit through the lateral cortex at a 90\u00b0 angle ~0.150\u2009mm below a gradient refractive index (GRIN) lens. We empirically derived a volume of interest (VOI)\u2009=\u20090.0161\u2009\u00b1\u20090.000675\u2009mm<jats:sup>3<\/jats:sup> during each imaging session by aggregating the Z\u2010stacks between the first (superior) and last (inferior) in\u2010focus LIMB slice. LIMB postimplantation revealed very limited bacteria detection at 1\u2009h, but by 3\u2009h,\u00a056.8% of the implant surface was covered by ECFP+ bacteria, and the rest were covered by GFP+ host cells. 3D volumetric rendering of the GFP+ and ECFP+ or RFP+ voxels demonstrated exponential MRSA growth between 3 and 6\u2009h\u00a0in the Z\u2010plane, which was validated with cross\u2010sectional ex vivo bacterial burden analyses demonstrating significant growth by ~2\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> CFU\/h\u00a0on the implant from 2 to 12\u2009h\u00a0post\u2010op (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05; <jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u2009&gt;\u20090.98). Collectively, these results show the competition at the surface is completed by 3\u2009h\u00a0in this model and demonstrate the potential of LIMB to elucidate mechanisms of bacterial colonization, the host immune response, and the efficacy of antimicrobials.<\/jats:p>","DOI":"10.1002\/jor.25716","type":"journal-article","created":{"date-parts":[[2023,10,9]],"date-time":"2023-10-09T18:49:58Z","timestamp":1696877398000},"page":"531-538","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Longitudinal intravital imaging of the bone marrow\u00a0for analysis of the race for the surface in a murine osteomyelitis model"],"prefix":"10.1002","volume":"42","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6197-2506","authenticated-orcid":false,"given":"Chao","family":"Xie","sequence":"first","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"},{"name":"Department of Orthopaedics and Rehabilitation University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youliang","family":"Ren","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"},{"name":"Department of Orthopaedics and Rehabilitation University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason","family":"Weeks","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"},{"name":"Department of Orthopaedics and Rehabilitation University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua","family":"Rainbolt","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Howard M.","family":"Kenney","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Xue","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faith","family":"Allen","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ye","family":"Shu","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allie J. H.","family":"Tay","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6638-9029","authenticated-orcid":false,"given":"Sashank","family":"Lekkala","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu\u2010Chi A.","family":"Yeh","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"},{"name":"Department of Orthopaedics and Rehabilitation University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7521-6154","authenticated-orcid":false,"given":"Gowrishankar","family":"Muthukrishnan","sequence":"additional","affiliation":[{"name":"Center for Musculoskeletal Research University of Rochester Medical Center Rochester New York USA"},{"name":"Department of Orthopaedics and Rehabilitation University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ann L.","family":"Gill","sequence":"additional","affiliation":[{"name":"Department of Microbiology and\u00a0Immunology University of Rochester Medical Center Rochester New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven R.","family":"Gill","sequence":"additional","affiliation":[{"name":"Department of Microbiology and\u00a0Immunology University of Rochester Medical Center Rochester New York 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