title: Deep vascular imaging in wounds by two-photon fluorescence microscopy.
abstract: Deep imaging within tissue (over 300 μm) at micrometer resolution has become possible with the advent of two-photon fluorescence microscopy (2PFM). The advantages of 2PFM have been used to interrogate endogenous and exogenous fluorophores in the skin. Herein, we employed the integrin (cell-adhesion proteins expressed by invading angiogenic blood vessels) targeting characteristics of a two-photon absorbing fluorescent probe to image new vasculature and fibroblasts up to ≈ 1600 μm within wound (neodermis)/granulation tissue in lesions made on the skin of mice. Reconstruction revealed three dimensional (3D) architecture of the vascular plexus forming at the regenerating wound tissue and the presence of a fibroblast bed surrounding the capillaries. Biologically crucial events, such as angiogenesis for wound healing, may be illustrated and analyzed in 3D on the whole organ level, providing novel tools for biomedical applications.
authors.affiliation: Department of Chemistry, University of Central Florida, Orlando, Florida, USA.
authors.forename: Ciceron O
authors.initials: CO
authors.lastname: Yanez
authors.forename: Alma R
authors.initials: AR
authors.lastname: Morales
authors.forename: Xiling
authors.initials: X
authors.lastname: Yue
authors.forename: Takeo
authors.initials: T
authors.lastname: Urakami
authors.forename: Masanobu
authors.initials: M
authors.lastname: Komatsu
authors.forename: Tero A H
authors.initials: TA
authors.lastname: Järvinen
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Belfield
mesh_terms: D000818:Animals; D002196:Capillaries / Q000293:injuries / Q000648:ultrastructure*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D005456:Fluorescent Dyes; D006097:Granulation Tissue / Q000293:injuries / Q000648:ultrastructure*; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods*; D016023:Integrins / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D036641:Microscopy, Fluorescence, Multiphoton; D018919:Neovascularization, Physiologic; D012867:Skin / Q000098:blood supply* / Q000293:injuries / Q000648:ultrastructure*; D014945:Wound Healing / Q000502:physiology; D014950:Wounds, Penetrating / Q000473:pathology
chemical_list: D005456:Fluorescent Dyes; D016023:Integrins