Institute of Laser and Optoelectronics Technology, Fujian Provincial Key Laboratory for Photonics Technology, Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, China.
Skin Res Technol. 2009 Nov;15(4):489-95. doi: 10.1111/j.1600-0846.2009.00395.x.
BACKGROUND/PURPOSE: A real-time, non-invasive method will confer a benefit for the diagnosis and treatment of localized scleroderma (LS) in the clinic. The aim of this work was to demonstrate the potential of multiphoton laser scanning microscopy (MPLSM) for diagnosing LS and monitoring the treatment response in vivo.
Three sclerodermatous skin specimens and two normal skin specimens were investigated using MPLSM based on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG). MPLSM consists of a femtosecond Ti:sapphire laser and a scanning inverted microscope. Several parameters such as the epidermal thickness, the orientation ratio index of collagen bundles (ORICB), the spacing of collagen fibrin as well as the SHG to TPEF index of the dermis (STID) were developed to quantitatively discriminate the sclerodermatous skin from the normal skin.
The morphological differences were visualized obviously in the TPEF/SHG images of human skin (normal and sclerodermatous). The values of the developed parameters in normal skin were significantly different from that in sclerodermatous skin (P<0.05).
MPLSM could discriminate the sclerodermatous skin from the normal skin. With the advent of the clinical portability of typical MPLSM, this technique has great potential for application in the in vivo diagnosis of LS as well as for monitoring the treatment response.
背景/目的：一种实时、非侵入性的方法将有益于局部性硬皮病（LS）的临床诊断和治疗。本研究旨在展示多光子激光扫描显微镜（MPLSM）在诊断 LS 以及在体监测治疗反应方面的潜力。
使用基于双光子激发荧光（TPEF）和二次谐波产生（SHG）的 MPLSM 对 3 例硬皮病皮肤标本和 2 例正常皮肤标本进行了研究。MPLSM 由飞秒钛宝石激光和扫描倒置显微镜组成。开发了几个参数，如表皮厚度、胶原束取向比指数（ORICB）、胶原纤维间距以及真皮的 SHG 与 TPEF 指数（STID），以定量区分硬皮病皮肤与正常皮肤。
人皮肤（正常和硬皮病）的 TPEF/SHG 图像中明显显示出形态学差异。正常皮肤的参数值与硬皮病皮肤的参数值有显著差异（P<0.05）。
MPLSM 可以区分硬皮病皮肤和正常皮肤。随着典型 MPLSM 的临床便携性的出现，这项技术在 LS 的体内诊断以及治疗反应监测方面具有很大的应用潜力。
