Dendritic cells (DCs), macrophages (Mf), and T cells are major components of the skin immune system, but their interstitial spatial organization is poorly characterized. Using four-channel whole-mount immunofluorescence staining of the human dermis, we demonstrated the three-dimensional distribution of CD31þ blood capillaries, LYVE-1þ lymphatics, discrete populations of CD11cþ myeloid DCs, FXIIIaþ Mf, and lymphocytes. We showed phenotypic and morphological differences in situ between DCs and Mf. DCs formed the first dermal cellular layer (0–20 mm beneath the dermoepidermal junction), Mf were located deeper (40–60 mm), and CD3þ lymphocytes were observed throughout (0–60 mm). Below this level, DCs, T cells, and the majority of Mf formed stable perivascular sheaths. Whole-mount imaging revealed the true extent of dermal leukocytes previously underestimated from cross-section views. The total area of apical dermis (0–30 mm) contained approximately 10-fold more myeloid DCs than the entire blood volume of an average individual. Surprisingly, o1% of dermal DCs occupied lymphatics in freshly isolated skin. Dermal DCs rapidly accumulated within lymphatics, but Mf remained fixed in skin explants cultured ex vivo. The leukocyte architecture observed in normal skin was distorted in inflammation and disease. These studies illustrate the micro-anatomy of dermal leukocytes and provide further insights into their functional organization.