Multiphoton laser scanning microscopy of localized scleroderma.
OBJECTIVE
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.


METHODS
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.


RESULTS
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).


CONCLUSIONS
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.
Lu
Kecheng
K
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.
Chen
Jianxin
J
Zhuo
Shuangmu
S
Zheng
Liqin
L
Jiang
Xingshan
X
Zhu
Xiaoqin
X
Zhao
Jingjun
J
D000328:Adult; D000368:Aged; D003094:Collagen / Q000378:metabolism; D046169:Dermoscopy / Q000295:instrumentation / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods; D004548:Elasticity; D004817:Epidermis / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005583:Fourier Analysis; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D012594:Scleroderma, Localized / Q000473:pathology*
D003094:Collagen