Herein, we integrate cell-imprinted substrate (CIS) and allochroic-graphene oxide (AGO) for specific visualization sorting of hepatocellular carcinoma cells. The state-of-the-art-of detection method relies on the enzyme linked immunosorbent assay (ELISA)-like sandwich strategy with hierarchical recognition. The target tumor cells are first selectively captured by the CIS based on cell imprinted recognition, and then specifically labeled with AGO by boronate affinity recognition between boronic acid on AGO and cis-diols on the surface of target cells. The selectively recognition of CIS for target template cells is verified by cell function experiments. It is also worth mentioning that the AGO can specifically recognize target tumor cells under physiological pH, and then perform signal amplification and output through pH-triggered allochroism. The CIS linked AGO for cell assay (CIS-AGO-CA) is successfully used for visualization detection of human hepatocarcinoma HLE cells from hepatocyte suspension. When the hepatocyte suspension is spiked with 1.0 × 10⁵ cells, the recoveries of CIS-AGO-CA are 80.67 ± 4.33% for target HLE cells, and only 12.00 ± 1.00% for non-target Hep3B cells. It is worth emphasizing that the CIS-AGO-CA process is antibody-free. Therefore, this novel ELISA-like sandwich strategy is high specificity, cost-efficient and easy-to-use, and exhibits great prospect in the visualization sorting of tumor subpopulation.

本文中，我们整合了细胞印迹底物（CIS）和异色石墨烯氧化物（AGO），以对肝细胞癌细胞进行特定的可视化排序。最新的检测方法依赖于具有层次识别功能的类似酶联免疫吸附测定（ELISA）的三明治策略。首先基于细胞印迹识别，由CIS选择性捕获靶肿瘤细胞，然后通过AGO上的硼酸和顺式之间的硼酸酯亲和力识别，用AGO特异性标记目标肿瘤细胞。-靶细胞表面上的二醇。通过细胞功能实验验证了对靶模板细胞的CIS选择性识别。还值得一提的是，AGO可以在生理pH下特异性识别靶肿瘤细胞，然后通过pH触发的异色作用进行信号放大和输出。CIS连接的用于细胞测定的AGO（CIS-AGO-CA）已成功用于从肝细胞悬液中可视化检测人肝癌HLE细胞。当肝细胞悬液掺入1.0×10 ^5时对于目标HLE细胞，CIS-AGO-CA的回收率为80.67±4.33％，对于非目标Hep3B细胞则仅为12.00±1.00％。值得强调的是，CIS-AGO-CA过程不含抗体。因此，这种新颖的类似ELISA的夹心策略具有高特异性，高成本效益和易于使用的特点，在肿瘤亚群的可视化分类中具有广阔的前景。