title: Whole-mount imaging of the mouse hindlimb vasculature using the lipophilic carbocyanine dye DiI.
abstract: The availability of transgenic disease backgrounds and the accessibility of molecular research reagents have contributed to make the mouse ischemic hindlimb the model of choice for many studies of angiogenesis, and to investigate new treatments for peripheral artery disease. A limitation of these models involves our inability to easily visualize the regenerated vascular architecture. Approaches such as micro-computed tomography and micro-angiography are expensive, technically demanding and not available to many laboratories. Here we describe a rapid and inexpensive adaptation of a vascular staining procedure for precise imaging of the mouse hindlimb vasculature. We introduced two technical modifications and an analytical extension to the original method including (i) pre-skinning of the muscle prior to fixation that preserves tissue integrity, (ii) mild pressure-desiccation subsequent to fixing that enhances resolution and image penetration, and (iii) reconstruction of confocal data into 3D images. The procedure provides resolution that is equivalent or superior to other approaches at a fraction of the cost, time and technology required.
authors: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA.|Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Jeffrey
authors: J
authors: Boden
authors: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Jianqin
authors: J
authors: Wei
authors: Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: George
authors: G
authors: McNamara
authors: Department of Bioengineering, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Hans
authors: H
authors: Layman
authors: Diabetes Research Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Midhat
authors: M
authors: Abdulreda
authors: Department of Bioengineering, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Fotios
authors: F
authors: Andreopoulos
authors: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, Miami, FL, USA.|Vascular Biology Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
authors: Keith A
authors: KA
authors: Webster
mesh_terms: D000818:Animals; D002232:Carbocyanines / Q000008:administration & dosage / Q000737:chemistry*; D006614:Hindlimb / Q000098:blood supply* / Q000293:injuries / Q000473:pathology; D021621:Imaging, Three-Dimensional; D007511:Ischemia / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal / Q000379:methods*
chemical_list: D002232:Carbocyanines; C024286:3,3'-dioctadecylindocarbocyanine
keywords: DiI; hindlimb; imaging; ischemia; perfusion; vascular