细胞膜表面蛋白质糖基化是一种广泛存在、结构复杂多变的蛋白质翻译后修饰，在细胞和机体内发挥着重要功能，其变化参与调节细胞识别、运动迁移、免疫应答、物质运输、细胞分化、信号转导等重要生物学过程。其功能紊乱会导致炎症、肿瘤等疾病的发生。为此，发展高分辨的精准测量技术可以获取细胞膜表面糖蛋白动态修饰过程中的空间分布波动信息，从而可对细胞病理变化过程进行深入解析，具有重要的理论和实践意义。本项目提出利用核酸适配体及生物正交反应等对糖蛋白进行特异性识别和编码式标记，结合短链核酸DNA分子动态杂交诱导的纳米级成像点积累技术（DNA-PAINT），在细胞体系突破光学衍射极限，克服细胞背景干扰，实现具有单分子灵敏度的超高分辨膜糖蛋白成像，特别是多种代表性糖分子及蛋白多重成像测量（如岩藻糖、唾液酸以及与细胞癌变关联的转铁蛋白，与耐药性关联的p-糖蛋白等），进而揭示生物大分子动态修饰的特征和调控机制。

Protein glycosylation on cell membrane, one of the widespread post-translational modifications with complicated and variety structures, plays important roles in cell and organism. The changes of protein glycosylation participate in a series of important biological processes, including cell recognition, migration, differentiation, immune response, nutrient transportation, signal transduction and so on, which might induce malignant disease such as inflammation and tumor. Therefore, the development of novel detection techniques with high spatial resolution and specificity is fundamentally significant, which can capture the dynamic modification process on the cell membrane and reveal the cellular pathological changes. This proposal is aimed to break through the optical diffraction limit and background interference from the cell and achieve ultrasensitive and super-resolution imaging for membrane glycoproteins, and especially for the multiplex imaging measurement of representative glycans and proteins, including fucose, sialic acid, transferrin (which is related to the cell cancerous progression) and p-glycoprotein (which is related to the drug resistance properties). The super-resolution imaging techniques will be carried out based on the DNA points accumulation for imaging in nanoscale topography (DNA-PAINT) induced by the dynamic hybridization of short DNA chains. The dynamic and high resolution messages will provide new insight into the elucidation of detailed functionality of glycoproteins on the cell membrane.