title: Analysis of Coronary Vessels in Cleared Embryonic Hearts.
abstract: 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.
authors.affiliation: Developmental Biology of Birth Defects, UCL Institute of Child Health; s.ivins@ucl.ac.uk.
authors.forename: Sarah
authors.initials: S
authors.lastname: Ivins
authors.affiliation: Developmental Biology of Birth Defects, UCL Institute of Child Health.
authors.forename: Catherine
authors.initials: C
authors.lastname: Roberts
authors.affiliation: MRC Centre for Regenerative Medicine, SCRM Building, University of Edinburgh.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Vernay
authors.affiliation: Developmental Biology of Birth Defects, UCL Institute of Child Health.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Scambler
mesh_terms: D000818:Animals; D001565:Benzoates; D019905:Benzyl Alcohol; D003331:Coronary Vessels / Q000000981:diagnostic imaging*; D006321:Heart / Q000000981:diagnostic imaging* / Q000196:embryology*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D018613:Microscopy, Confocal; D012997:Solvents
chemical_list: D001565:Benzoates; D012997:Solvents; D019905:Benzyl Alcohol; C006723:benzyl benzoate