Anaesthetized embryos at indicated stage were mounted in 1.0% low-melt agarose (Sigma Aldrich) for imaging19 (link). Images were acquired by using an Olympus FluoView FV1000 laser scanning confocal microscope. Z-stacks were acquired with a 3 μM z-step, and images were 3D rendered. Neurolucida software (MBF Bioscience) was used to analyze the length of ISVs and SIV.
Protocol detail
Imaging Anesthetized Embryonic Vasculature
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Agarose
Embryos
Laser scanning microscope
Corresponding Organization :
Other organizations : Houston Methodist
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Variable analysis
independent variables
- Anaesthetized embryos at indicated stage
- Length of ISVs (intersegmental vessels)
- Length of SIV (subintestinal vessel)
- Mounting of anaesthetized embryos in 1.0% low-melt agarose (Sigma Aldrich) for imaging
- Imaging using an Olympus FluoView FV1000 laser scanning confocal microscope
- Acquisition of z-stacks with a 3 μM z-step and 3D rendering of images
- Analysis of ISVs and SIV lengths using Neurolucida software (MBF Bioscience)
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