Institute of Photonic Technology Jena, Albert-Einstein-Straße 9, 07745, Jena, Germany.
Br J Dermatol. 2013 Oct;169(4):794-803. doi: 10.1111/bjd.12427.
The combination of coherent anti-Stokes Raman scattering (CARS), second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) imaging--referred to as multimodal imaging--provides complementary contrast based on molecular vibrations, the structure of various tissue components and endogenous fluorophores, respectively.
To present a comprehensive overview of the appearance of human skin in multimodal imaging.
Multimodal imaging of unstained skin cross-sections of 32 individuals was performed using a laser scanning microscope and picosecond laser pulse for excitation.
The epidermis, dermis and subcutis are distinguishable in all three applied modalities, but are unveiled best in multimodal images. While the subcutis is dominated by the CARS signal, predominately SHG and the secondary TPEF signal detect the dermis. In contrast, no SHG signal is detected in the epidermis, whereas CARS and TPEF show equal contributions. Additionally, the appearance of the major skin appendages is described, i.e. the hair follicle, sebaceous and sweat glands, and blood vessels belonging to the vascular system. All four investigated functional units show a characteristic morphochemistry in TPEF and CARS, allowing identification of further subunits, e.g. the major components of the hair follicle, while the SHG signal delineates the localization of the functional units.
Multimodal imaging is a powerful tool to investigate human skin by providing high contrast based on the molecular constitution. It is therefore suggested that multimodal imaging has a high potential in application to dermatological research and clinical diagnostics of various skin alterations.
相干反斯托克斯拉曼散射(CARS)、二次谐波产生(SHG)和双光子激发荧光(TPEF)成像的组合——称为多模态成像——分别基于分子振动、各种组织成分的结构和内源性荧光团提供互补对比。
全面介绍多模态成像中人体皮肤的外观。
使用激光扫描显微镜和皮秒激光脉冲对 32 个人体未染色皮肤切片进行多模态成像。
在所有三种应用模式中,表皮、真皮和皮下组织均可分辨,但在多模态图像中最佳显现。虽然皮下组织主要由 CARS 信号主导,但主要的 SHG 和次要的 TPEF 信号检测真皮。相比之下,表皮中未检测到 SHG 信号,而 CARS 和 TPEF 显示出相等的贡献。此外,还描述了主要皮肤附属物的外观,即毛囊、皮脂腺和汗腺以及属于脉管系统的血管。在 TPEF 和 CARS 中,所有四个研究的功能单元均显示出特征形态化学,从而可以识别出进一步的亚单位,例如毛囊的主要成分,而 SHG 信号描绘了功能单元的定位。
多模态成像通过基于分子构成提供高对比度,是研究人体皮肤的有力工具。因此,建议多模态成像在皮肤科研究和各种皮肤改变的临床诊断中有很高的应用潜力。