title: Optimization of Vascular Casting for Three-Dimensional Fluorescence Cryo-Imaging of Collateral Vessels in the Ischemic Rat Hindlimb.
abstract: Development of collateral vessels, arteriogenesis, may protect against tissue ischemia, however, quantitative data on this process remain scarce. We have developed a technique for replicating the entire arterial network of ischemic rat hindlimbs in three dimensions (3D) based on vascular casting and automated sequential cryo-imaging. Various dilutions of Batson's No. 17 with methyl methacrylate were evaluated in healthy rats, with further protocol optimization in ischemic rats. Penetration of the resin into the vascular network greatly depended on dilution; the total length of casted vessels below 75 µm was 13-fold higher at 50% dilution compared with the 10% dilution. Dilutions of 25-30%, with transient clamping of the healthy iliac artery, were optimal for imaging the arterial network in unilateral ischemia. This protocol completely filled the lumina of small arterioles and collateral vessels. These appeared as thin anastomoses in healthy legs and increasingly larger vessels during ligation (median diameter 1 week: 63 µm, 4 weeks: 127 µm). The presented combination of quality casts with high-resolution cryo-imaging enables automated, detailed 3D analysis of collateral adaptation, which furthermore can be combined with co-registered 3D distributions of fluorescent molecular imaging markers reflecting biological activity or perfusion.
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Janina C V
authors.initials: JC
authors.lastname: Schwarz
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Monique G J T B
authors.initials: MG
authors.lastname: van Lier
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Erik N T P
authors.initials: EN
authors.lastname: Bakker
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Judith
authors.initials: J
authors.lastname: de Vos
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Jos A E
authors.initials: JA
authors.lastname: Spaan
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Ed
authors.initials: E
authors.lastname: VanBavel
authors.affiliation: Department of Biomedical Engineering and Physics,Academic Medical Center,University of Amsterdam,Meibergdreef 9, 1105 AZ Amsterdam,The Netherlands.
authors.forename: Maria
authors.initials: M
authors.lastname: Siebes
mesh_terms: D000818:Animals; D001160:Arterioles / Q000000981:diagnostic imaging / Q000473:pathology; D001808:Blood Vessels / Q000000981:diagnostic imaging* / Q000473:pathology*; D016319:Corrosion Casting; D004853:Epoxy Resins; D006614:Hindlimb / Q000098:blood supply* / Q000000981:diagnostic imaging*; D006652:Histological Techniques / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007511:Ischemia / Q000000981:diagnostic imaging* / Q000473:pathology*; D008026:Ligation; D008297:Male; D008768:Methylmethacrylates / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004853:Epoxy Resins; D008768:Methylmethacrylates
keywords: 3D imaging; collateral vessels; hindlimb ischemia; vascular casting; vascular imaging