Department of Immunology, Weizmann Institute of Science, Rehovot, Israel.
Methods Mol Biol. 2022;2380:3-13. doi: 10.1007/978-1-0716-1736-6_1.
Long-lasting immunity depends on generation of antibody forming cells in germinal centers (GCs). Conventional methods such as immunohistology and intravital live imaging have been used extensively to investigate the location of cellular assemblies within tissues as well as their dynamic motility and cellular interactions. Two photon laser scanning microscopy (TPLSM) intravital imaging allows scanning of large areas within tissues and reveals multiple immune cell niches. Nonetheless, this type of imaging is limited by the depth of penetration and cannot capture effectively all of the GC niches within lymphoid organs. Here we describe a method to visualize antigen-specific T and B cells in multiple microanatomical locations and niches at the level of a whole organ. This large-scale imaging approach can greatly increase our understanding of the spatial distribution of immune cells and help obtain detailed 3D maps of their locations and quantities.
长期免疫依赖于生发中心（GCs）中抗体形成细胞的产生。免疫组织化学和活体成像等传统方法已被广泛用于研究组织内细胞组合的位置及其动态运动和细胞相互作用。双光子激光扫描显微镜（TPLSM）活体成像允许对组织内的大片区域进行扫描，并揭示多个免疫细胞龛。尽管如此，这种类型的成像受到穿透深度的限制，无法有效地捕获淋巴器官内所有的 GC 龛位。在这里，我们描述了一种在整个器官水平上可视化多个微解剖位置和龛位的抗原特异性 T 和 B 细胞的方法。这种大规模成像方法可以大大提高我们对免疫细胞空间分布的理解，并有助于获得其位置和数量的详细三维图谱。
