Twenty images from mid-Matrigel H & E sections of all animals in vehicle and propranolol groups were taken randomly at 40× (Zeiss Axiophot II, equipped with AxioCam MRc5 and supplemented with AxioVision Rel. 4.8 software). Luminal structures containing at least one RBC were counted as one vessel; the average from 20 images was expressed as vessels/mm2. Formalin-fixed, paraffin-embedded mid-Matrigel sections were processed and stained with mouse anti-human CD31 (1:40, Dako M0823) or mouse IgG (1:40, Dako X0931) as described18 (link).
Protocol detail
Quantifying Angiogenesis in Matrigel
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Variable analysis
independent variables
- Treatment group (vehicle or propranolol)
- Vessel density (vessels/mm^2)
- Matrigel sections
- Tissue processing and staining (Formalin-fixed, paraffin-embedded mid-Matrigel sections, stained with mouse anti-human CD31 or mouse IgG)
- Microscopy settings (40× magnification, Zeiss Axiophot II, AxioCam MRc5, AxioVision Rel. 4.8 software)
- Positive control: Mouse anti-human CD31 staining
- Negative control: Mouse IgG staining
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