Human skin harbors various immune cells that are crucial for the control of injury and infection. However, the current understanding of immune cell function within viable human skin tissue is limited. We developed an ex vivo imaging approach in which fresh skin biopsies are mounted and then labeled with nanobodies or antibodies against cell surface markers on tissue-resident memory CD8⁺ T cells, other immune cells of interest, or extracellular tissue components. Subsequent longitudinal imaging allows one to describe the dynamic behavior of human skin-resident cells in situ. In addition, this strategy can be used to study immune cell function in murine skin. The ability to follow the spatiotemporal behavior of CD8⁺ T cells and other immune cells in skin, including their response to immune stimuli, provides a platform to investigate physiological immune cell behavior and immune cell behavior in skin diseases. The mounting, staining and imaging of skin samples requires ~1.5 d, and subsequent tracking analysis requires a minimum of 1 d. The optional production of fluorescently labeled nanobodies takes ~5 d.

人的皮肤具有各种免疫细胞，这些免疫细胞对于控制损伤和感染至关重要。但是，目前对人类皮肤组织中免疫细胞功能的了解有限。我们开发了一种离体成像方法，其中安装了新鲜的皮肤活检样品，然后用纳米抗体或针对组织驻留记忆CD8 ⁺ T细胞，其他感兴趣的免疫细胞或细胞外组织成分上的细胞表面标志物的抗体标记。随后的纵向成像使人们能够原位描述人类皮肤驻留细胞的动态行为。另外，该策略可用于研究鼠皮中的免疫细胞功能。遵循CD8 ⁺时空行为的能力皮肤中的T细胞和其他免疫细胞，包括它们对免疫刺激的反应，为研究生理性免疫细胞行为和皮肤疾病中的免疫细胞行为提供了一个平台。皮肤样品的安装，染色和成像需要约1.5 d，随后的跟踪分析至少需要1 d。荧光标记的纳米抗体的可选生产需要约5 d。