Multiphoton Microscopic Imaging Of Sclerosing Atrophic And Scleroderma

Objective:To investigate the feasibility of the multiphoton laser scanning microscopy as a new,non-invasive and easy diagnosis and identification method for sclerosing atrophic and scleroderma.Content:Multi-photon microscopic imaging technology has characteristics of high resolution,deep penetration depth,two-dimensional imaging and so on which can observe the human skin lesions in vivo,noninvasive,dynamic and repetitive.Human endogenous signals are used to for its imaging.For example,human keratin,cytoplasm,nucleolus,elastic fibers,collagen fibers and intercellular matrix show red images,representing TPEF signal,collagen fibers show green images,representing SHG signal and the nucleus,vascular lumen does not send fluorescent signal.In this study,TPEF/SHG images can clearly show the skin stratum corneum,epidermis,dermal papillae,capillaries,collagen fibers and elastic fibers and other tissue images which can be easier for clinical physicians to diagnose skin diseases,estimate the conditions and guide the treatment.Considering the advantages of this technique in the basic diagnosis of dermatology and the basic research of histomorphology,this paper applies multi-photon microscopic imaging technology to the diagnosis and identification of two common skin diseases sclerosing atrophic moss and scleroderma.Methods: In this study,Multiphoton laser scanning microscopy was used to image the diseased tissue of sclerosing atrophic moss and scleroderma.High resolution and high contrast TPEF/SHG images were obtained to describe the tissue characteristics and difference of the two skin diseases.Meanwhile,we also defined the Orientation Index of Collagen Bundles(OICB),and the OICB value was calculated by Image J software through FFT conversion to quantitatively assess the difference in tissue morphology.Results: According to TPEF/SHG images,in the histomorphological,sclerosing atrophic moss lesions skin can be seen vacuolar basal cells,hippocampal suppository,subcutaneous bullae and other structures.There are many fibrous tissue fractures and broken in the shallow layer of dermis,and a large number of helical collagen fibers and wave-like elastic fibers are arranged in parallel to each other in the deep layer of dermis.Scleroderma lesions skin epidermis basal layer has a flat trend.Some dermal fibrous tissue fracture,broken,loss of normal linear or wave-like structure in the shallow layer of dermis.And in the deep layer of dermis elastic fiber undulating amplitude becomes smaller,collagen fibers were significantly thicker,they are arranged in parallel.In addition,when scleroderma OICB was compared with sclerosing atrophic OICB,in the shallow layer of dermis the statistic analysis showed a statistically significant difference in the data,but in the deep layer of dermis there was no statistically significant difference.Conclusion:The experimental results show that Multiphoton laser scanning microscopy can be applied to distinguish sclerosing atrophic moss and scleroderma in histomorphology,which will provide a laboratory basis for the noninvasive diagnosis and differential diagnosis of these two skin diseases in foreseeable future.