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Description

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.

Technical Details

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).

Biological Sources

NCBI Organism Classification
Mus musculus
(house mouse)
Cell Type
hematopoietic stem cell
Cellular Component
external side of plasma membrane
integral to membrane

Attribution

Names
Samir Taoudi
Alexander Medvinsky
Published
Proc Natl Acad Sci. 2007. 104: 9399-9403.
Link
http://intl.pnas.org/content/104/22/93...
Pubmed
17517650

Imaging

Image Type
recorded image
Imaging Mode
single-spot confocal microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
primary antibody plus labeled secondary antibody
R-phycoerythrin (PE)
Allophycocyanin (APC)
Processing History
unprocessed raw data
Data Qualifiers
raw, unprocessed data

Sample Preparation

Methods
fixed tissue
Relation To Intact Cell
whole mounted tissue

Dimensions

Spatial Axis Image Size Pixel Size
X 1027px ——
Y 892px ——
Z 150 1.0μm
*CIL – Cell Image Library accession number. Please use this to reference an image.