嵌合抗原受体（CAR）识别肿瘤相关抗原（TAA），可以有效地靶向肿瘤细胞，而无需使用人类主要组织相容性复合物（MHC）。但是，在临床实践中，CAR具有剂量确定不准确的问题，而可以解决这个问题的方法通常产生细胞毒性物质，例如绿色荧光蛋白（GFP）插入。因此，在本研究中，我们试图使用高度生物兼容的应力促进炔-叠氮环加成（SPAAC）将无害荧光标记锚定在CAR-T细胞膜上，而不产生任何副产品。我们的共轭荧光标记在CAR-T细胞表面上稳定至少两周，并具有出色的光学稳定性和测量学性能。此外，这种方法可以快速定量活性CAR-T细胞，而不影响它们的活性。因此，这种方法是一种有前途的可靠策略，用于准确诊断和治疗癌症。

Chimeric antigen receptors (CARs) recognizing tumor-associated antigens (TAAs) effectively target tumor cells without using the major histocompatibility complex (MHC). However, CARs have inaccurate dose determination in clinical practice, and the methods that can solve this problem often produce cytotoxic substances, such as green fluorescent protein (GFP) insertion. Therefore, in this study, we tried to anchor harmless fluorescent labels on CAR-T cell membranes using highly biologically compatible strain-promoted alkyne-azide cycloaddition (SPAAC) without any byproducts. Our conjugated fluorescent label was stable on the CAR-T cell surface for at least two weeks, with excellent light stability and metrology. Also, this method enabled the rapid quantification of the living CAR-T cells without affecting their activity. Thus, this method is a promising reliable strategy for accurately diagnosing and treating cancer.