Deep vascular imaging in wounds by two-photon fluorescence microscopy

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...

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Bibliographic Details
Published in:PloS one 2013-07, Vol.8 (7), p.e67559-e67559
Main Authors: Yanez, Ciceron O, Morales, Alma R, Yue, Xiling, Urakami, Takeo, Komatsu, Masanobu, Järvinen, Tero A H, Belfield, Kevin D
Format: Article
Language:English
Subjects:
Angiogenesis
Animals
Biology
Biomedical materials
Blood vessels
Bone surgery
Capillaries
Capillaries - injuries
Capillaries - ultrastructure
Chemical compounds
Chemistry
Fibroblasts
Fibroblasts - cytology
Fibroblasts - physiology
Fluorescence
Fluorescence microscopy
Fluorescent Dyes
Fluorophores
Granulation
Granulation Tissue - injuries
Granulation Tissue - ultrastructure
Imaging, Three-Dimensional - instrumentation
Imaging, Three-Dimensional - methods
Integrins
Integrins - metabolism
Lesions
Male
Medical imaging
Medical research
Medicine
Mice
Mice, Inbred BALB C
Microscopy
Microscopy, Fluorescence, Multiphoton
Neovascularization, Physiologic
Peptides
Proteins
Rodents
Skin
Skin - blood supply
Skin - injuries
Skin - ultrastructure
Wound healing
Wound Healing - physiology
Wounds and injuries
Wounds, Penetrating - pathology
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0067559