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Visualization and Analysis of Pharyngeal Arch Arteries using Whole-mount Immunohistochemistry and 3D Reconstruction
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Developmental Biology
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JoVE Journal Developmental Biology
Visualization and Analysis of Pharyngeal Arch Arteries using Whole-mount Immunohistochemistry and 3D Reconstruction

Visualization and Analysis of Pharyngeal Arch Arteries using Whole-mount Immunohistochemistry and 3D Reconstruction

DOI:
10.3791/60797-v

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10:02 min

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March 31, 2020

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1Department of Cell Biology and Molecular Medicine, New Jersey Medical School,Rutgers Biomedical and Health Sciences, 2Multidisciplinary PhD Program in Biomedical Sciences: Cell Biology, Neuroscience and Physiology Track, New Jersey Medical School,Rutgers Biomedical and Health Sciences

Chapters

  • 00:04Introduction
  • 00:55Embryo Staining and Embedding
  • 03:03Methanol Dehydration and Tissue Clearing
  • 04:04Embryo Mounting
  • 04:40Third Pharyngeal Arch (PA) Surfacing
  • 05:57Surfaced Structure Masking
  • 07:01Endothelial Cell Quantification
  • 08:19Results: Representative PA and PAA Surfacing and EC Quantification
  • 09:28Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a protocol to visualize and analyze the pharyngeal arch arteries 3, 4, and 6 of mouse embryos using whole-mount immunofluorescence, tissue clearing, confocal microscopy, and 3D reconstruction.

Tags

Keywords: VisualizationAnalysisPharyngeal Arch ArteriesWhole-mount Immunohistochemistry3D ReconstructionEndothelial Cell QuantificationAortic Arch ArteriesCongenital Heart DiseaseVessel FormationRemodelingLight-sheet MicroscopyMouse EmbryoFixationPermeabilizationBlocking BufferPrimary AntibodySecondary AntibodyPlastic Paraffin Molds

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