Deep Tissue Staining with anti-CD3 to Identify T-cells

Human tonsil stained with Biocare Medical BC 33 in a 1:100 concentration.

Formalin-fixed, paraffin-embedded human tonsil tissue ranging in size up to 3mm was de-waxed, pretreated, and subject to antigen retrieval using an EDTA/Methanol buffer system.  The tissue was stained using Biocare Medical anti-CD3 clone BC33 and a nuclear stain, TO-PRO-3 (Thermo-Fisher). The tissue was cleared using a hybrid iDISCO protocol before imaging on Alpenglow Biosciences Aurora™ 3Di Hybrid Open Top Light Sheet (HOTLS) fluorescent microscope. For more information and a list of optimized antibodies for deep tissue imaging, see our brochure “Staining with Immunomarkers within Deep Tissues”.