Multigated DNA Cascade Amplifier for Ultrasensitive Spatiotemporal Imaging of PLS3 mRNA at the Single-Cell Level for Early Detection of Breast Cancer Metastasis

nanoflowers to facilitate intracellular delivery, while intracellular glutathione (GSH) and external UV light enabled programmed probe release. The combination of PLS3 mRNA-triggered hybridization chain reaction (HCR) and APE1-mediated enzyme catalytic amplification leads to a notable increase in detection sensitivity, achieving a detection limit of 5.8 aM. The MDCA was applied for in vitro sensing of PLS3 mRNA in breast cancer cell lines at the single-cell level. More importantly, with the aid of machine learning models, the MDCA facilitates the visualization of breast cancer metastasis in clinical tissue samples, with an excellent accuracy of more than 92%. This method holds great promise for early diagnosis, evaluating metastasis risk, and advancing precision oncology.