Department of Chemistry, University of Central Florida, Orlando, Florida, USA.
PLoS One. 2013 Jul 2;8(7):e67559. doi: 10.1371/journal.pone.0067559. Print 2013.
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.
随着双光子荧光显微镜（2PFM）的出现，已经可以在组织内进行深度成像（超过 300μm），达到微米分辨率。2PFM 的优势已被用于探测皮肤中的内源性和外源性荧光团。在此，我们利用双光子吸收荧光探针的整合素（由侵袭性血管生成血管表达的细胞粘附蛋白）靶向特性，在小鼠皮肤损伤处的伤口（真皮/肉芽组织）内成像新的血管和成纤维细胞，深度可达 ≈ 1600μm。重建结果显示了在再生伤口组织中形成的血管丛的三维（3D）结构，以及围绕毛细血管的成纤维细胞床的存在。这样，就可以在整个器官水平上以 3D 形式展示和分析对伤口愈合至关重要的生物学事件，如血管生成，为生物医学应用提供了新的工具。
