Human paraffin-embedded spleen tissue slices were prepared with a standard protocol of deparaffinization and rehydration. Antigen retrieval was done with Citrate-Buffered 1X pH6.0 at 95°C for 40 minutes. Tissue was washed with PBS/0.05% Tween 20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/ml of Alexa Fluor® 647 anti-human CD66a/c/e (clone ASL-32) Antibody (red) and Alexa Fluor® 594 anti-human CD45RO (clone UCHL1) antibody (green) overnight at 4°C. The image was scanned with a 10X objective and stitched with MetaMorph® software.

Human colorectal adenocarcinoma cell line HT-29 was stained with CD66a/c/e (clone ASL-32) Alexa Fluor® 647 (filled histogram) or mouse IgG2b, κ Alexa Fluor® 647 isotype control (open histogram).