Movie shows restricted distribution of PECAM-1[high] CD45+ cells to the luminal surface of the aortic endothelium from an E11.5 mouse embryo. PECAM-1[high] cells are in red, CD45+ cells are in green and the PECAM-1[high] CD45+ cells, defined as hematopoietic stem cells, can be seen in yellow. PECAM-1[high] CD45+ cells are scattered on the floor of the luminal surface, but not in subendothelial mesenchymal layers. Z-stack through whole-mount preparation of ventral aorta (150 images captured at 1.0 micron optical intervals) can be turned to allow clear observation of PECAM-1[high] CD45+ cells. Movie is Supporting Information Movie 1 and images from Z-stack are included in Fig 4 from Taoudi et al.
The aorta was dissected from E10.5-E11.5 embryos and incubated with PE-conjugated anti-PECAM-1 (clone MEC 13.3; BD Biosciences, Franklin Lakes, NJ) and APC-conjugated anti-CD45 (clone 30-F11; BD Biosciences) monoclonal antibodies, mounted and imaged on an inverted confocal microscope (DM IRE2: Leica).
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