This dataset demonstrates the identification of memory T cells in formalin-fixed, paraffin-embedded (FFPE) human tonsil tissue using optimized staining and clearing workflows. Tissue samples up to 3 mm were de-waxed, pretreated, and underwent antigen retrieval with an EDTA/Methanol buffer system. They were then stained with BioLegend anti-CD45RO (clone UCHL1) and the nuclear dye TO-PRO-3 (Thermo Fisher).

The tissue was cleared using a hybrid iDISCO protocol, ensuring deep antibody penetration, and imaged with the Aurora™ 3Di Hybrid Open Top Light Sheet (HOTLS) fluorescent microscope. This workflow preserves tissue integrity while enabling true 3D histology of memory T cell localization within lymphoid tissue.

Through integration with 3Dm data management and 3Dai AI-powered segmentation, researchers can perform reproducible quantification of CD45RO-positive cell distribution and spatial relationships. Such high-content, volumetric analysis supports both immunology research and translational applications in immuno-oncology, where memory T cell activity plays a pivotal role in therapeutic outcomes.