Dynamic Decoration of DNA Scaffolds for High‐Resolution Cancer Cell Subtyping

ABSTRACT Precise imaging of cancer cells serves as the foundation for subtype analysis, significantly advancing the development of precision medicine. The unavoidable cellular internalization of fluorescent labels constrains the resolution and timeliness, presenting a significant obstacle. This study introduces a dynamic decorating strategy of DNA scaffolds that enables the execution of four distinct Boolean logic operations through self‐assembly and self‐disassembly. By integrating the in‐built molecular circuit, the proposed assay achieved signal‐amplified detection of low‐abundance nucleic acid inputs. In the application of cell imaging, inputs were labeled with aptamers to operate membrane‐confined self‐assembly or self‐disassembly of DNA scaffolds on the cell surface, enabling simultaneous identification of distinct subtypes of cancer cells with high fidelity. The intrinsic durability of DNA scaffolds successfully prevented cellular internalization for up to 300 min, boosting the long‐duration imaging. Moreover, the assay was capable of profiling a broad spectrum of cancer cell abundances from as low as 0.1% to 10% in clinical blood samples, consistently achieving recognition efficiency exceeding 60%. These findings underscore the transformational potential of DNA scaffold‐based imaging tools in biological research and precision medicine.