Live imaging of skin immunity using two-photon microscopy: a short review

Two-photon excitation microscopy has become an important technique in dermatologic research, providing high-resolution imaging of living skin and its immune responses. Unlike conventional histology, two-photon microscopy allows direct observation of living tissues in real time at cellular resolution. This method enables the visualization of keratinocyte organization, skin appendages such as hair follicles and sebaceous glands, and the vascular, lymphatic, and neural networks within the dermis. It has also revealed dynamic immune processes in the skin. Clinical application is still limited by safety concerns and the high cost of equipment. Nonetheless, strategies based on autofluorescence, melanin scattering, and second harmonic generation (SHG) have been explored to visualize collagen and fibrosis in the dermis. This review summarizes the structural, functional, and translational aspects of two-photon skin imaging. We outline the technical principles, applications in animal and human studies, and potential implications for dermatologic research and clinical diagnosis.