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Two\u2010channel two\u2010photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta\u2010based macrophages using plasmonic gold nanorose as an imaging contrast agent.<\/jats:p><\/jats:sec><jats:sec><jats:title>Study Design\/Materials and Methods<\/jats:title><jats:p>Nanorose uptake by macrophages was identified by TPL microscopy in macrophage cell culture. <jats:italic>Ex vivo<\/jats:italic> aorta segments (8\u2009\u00d7\u20098\u2009\u00d7\u20092\u2009mm<jats:sup>3<\/jats:sup>) rich in macrophages from a rabbit model of aorta inflammation were imaged by TPL microscopy in combination with OCT. Aorta histological sections (5\u2009\u00b5m in thickness) were also imaged by TPL microscopy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Merged two\u2010channel TPL images showed the lateral and depth distribution of nanorose\u2010loaded macrophages (confirmed by RAM\u201011 stain) and other aorta components (e.g., elastin fiber and lipid droplet), suggesting that nanorose\u2010loaded macrophages are diffusively distributed and mostly detected superficially within 20\u2009\u00b5m from the luminal surface of the aorta. Moreover, OCT images depicted detailed surface structure of the diseased aorta.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Results suggest that TPL microscopy combined with OCT can simultaneously reveal macrophage distribution with respect to aorta surface structure, which has the potential to detect vulnerable plaques and monitor plaque\u2010based macrophages overtime during cardiovascular interventions. 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