Analysis of Coronary Vessels in Cleared Embryonic Hearts

Sarah Ivins, Catherine Roberts, Bertrand Vernay, Peter J Scambler

Research output: Contribution to journalArticlepeer-review


Whole mount visualization of the embryonic coronary plexus from which the capillary and arterial networks will form is rendered problematic using standard microscopy techniques, due to the scattering of imaging light by the thick heart tissue, as these vessels are localized deep within the walls of the developing heart. As optical clearing of tissues using organic solvents such as BABB (1 part benzyl alcohol to 2 parts benzyl benzoate) has been shown to greatly improve the optical penetration depth that can be achieved, we combined clearance of whole, PECAM1-immunostained hearts, with laser-scanning confocal microscopy, in order to obtain high-resolution images of vessels throughout the entire heart. BABB clearance of embryonic hearts takes place rapidly and also acts to preserve the fluorescent signal for several weeks; in addition, samples can be imaged multiple times without loss of signal. This straightforward method is also applicable to imaging other types of blood vessels in whole embryos.

Original languageEnglish
Number of pages9
JournalJournal of Visualized Experiments (JoVE)
Issue number118
Publication statusPublished - 7 Dec 2016


  • Developmental Biology
  • coronary vessels
  • BABB
  • PECAM1
  • embryonic heart
  • wholemount immunofluorescence
  • optical clearing
  • confocal microscopy


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