发展单细胞甚至单分子水平生物标志物的精准诊断有助于病变细胞在可治愈阶段提供早期治疗方法,利用大数据单细胞分析策略实现对每一个生物个体信息的原位精准提取。以生物标志物碱性磷酸酶（ALP）为例，作为肝脏相关癌症疾病检测的重要指标，亟需发展单细胞原位的单分子生物信息的提取方法。基于电化学方法的生物纳米孔道单分子信息研究利用纳米空间限域效应，对单个分子具有超高时间、空间分辨能力，目前已经实现对单链DNA的单碱基信息的识别和提取。然而传统生物纳米孔道受限于装置，难以实现对单个细胞的单个生物分子信息原位提取。本项目拟发展单细胞内基于纳米孔道单分子瞬态时频信息提取方法，创新性地结合生物纳米孔道与微操装置，突破传统纳米孔道应用场景的局限性，构建单细胞原位基于生物纳米孔道的单分子生物信息检测装置，并利用频谱分析以及深度学习策略，构建单细胞单分子相互作用谱图，发展为精准诊疗与研究的单个生物分子信息的研究策略。

The development of accurate diagnosis of single-cell or even single-molecule biomarkers helps diseased cells to provide early treatment in the curable phase, using the big data single-cell analysis strategy to achieve accurate in-situ extraction of each individual bioinformation. Taking the biomarker alkaline phosphatase (ALP) as an example, as an important indicator for the detection of liver-related cancer diseases, it is urgent to develop a single-cell in situ single-molecule biological information extraction method. The single-molecule information research of biological nanopore based on electrochemical method utilizes the nano-space confinement effect to have ultra-high time and spatial resolution ability for single molecules. At present, single-base information recognition and extraction of single-stranded DNA has been realized. However, traditional biological nanopore approaches are limited by devices, making it difficult to achieve in situ extraction of individual biomolecule information for a single cell. This project intends to develop a single-cell based single-molecule transient time-frequency information extraction method in a single cell, innovatively combine biological nanopore and micro-manipulators, break through the traditional nanopore application limitation, and construct single-cell in situ based bio-nano The single-molecule biological information detection device of the channel, and using spectrum analysis and deep learning strategies, constructs a single-cell single-molecule interaction spectrum, and develops the extraction of individual biomolecule information for precise diagnosis and research of the disease in the curable stage.