E10.5 embryos were fixed, stained, and analyzed as described (33 (link), 63 (link)). Embryos were stained with biotinylated anti–PECAM-1 (BD Biosciences, 553371) and anti–c-Kit (BD Biosciences, 553352) antibodies. Samples were mounted in a 1:2 mix of benzyl alcohol (Sigma-Aldrich, 402834) and benzyl benzoate (Acros Organics, 105862500) to increase tissue transparency and visualized with a Nikon A1RS confocal microscope. Three-dimensional reconstructions were generated from Z-stacks (80 to 200 optical sections) using Fiji software.
Protocol detail
Embryonic Angiogenesis and Hematopoiesis Imaging
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Antibodies
Benzyl alcohol
Benzyl benzoate
Confocal microscope
Embryos
Pecam 1
Reconstructions
Sections
Tissue
Corresponding Organization : University of Wisconsin Carbone Cancer Center
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Variable analysis
independent variables
- None explicitly mentioned
- Embryonic development and tissue structure
- Embryos were fixed, stained, and analyzed as described in references 33 and 63
- Embryos were stained with biotinylated anti–PECAM-1 and anti–c-Kit antibodies
- Samples were mounted in a 1:2 mix of benzyl alcohol and benzyl benzoate to increase tissue transparency
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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