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JoVE Journal

Developmental Biology

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

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10:02 min

March 31st, 2020

March 31st, 2020

9,375 Views

0:04

Introduction

0:55

Embryo Staining and Embedding

3:03

Methanol Dehydration and Tissue Clearing

4:04

Embryo Mounting

4:40

Third Pharyngeal Arch (PA) Surfacing

5:57

Surfaced Structure Masking

7:01

Endothelial Cell Quantification

8:19

Results: Representative PA and PAA Surfacing and EC Quantification

9:28

Conclusion

Transcript

This method allows each pharyngeal arch to be compartmentalized. Endothelial cell numbers can then be quantified in each arch as a means to study pharyngeal arch artery development. The main advantages of this approach are the ability to visualize

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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.

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