HIGHLIGHTS• A one-pot method was developed for the preparation of fluorescent mesoporous silica nanoparticles-carbon dots (MSNs-CDs) nanohybrid.• The MSNs-CDs nanohybrid showed stable and bright yellow emission, excellent biocompatibility, and specific targeting capability toward folate receptor-overexpressing cancer cells and can be applied as fluorescence imaging-guided drug carriers for effectively delivering anticancer drugs to tumor sites.ABSTRACT Multifunctional nanocarrier-based theranostics is supposed to overcome some key problems in cancer treatment. In this work, a novel method for the preparation of a fluorescent mesoporous silica-carbon dot nanohybrid was developed. Carbon dots (CDs), from folic acid as the raw material, were prepared in situ and anchored on the surface of amino-modified mesoporous silica nanoparticles (MSNs-NH 2 ) via a microwave-assisted solvothermal reaction. The as-prepared nanohybrid (designated MSNs-CDs) not only exhibited strong and stable yellow emission but also preserved the unique features of MSNs (e.g., mesoporous structure, large specific surface area, and good biocompatibility), demonstrating a potential capability for fluorescence imagingguided drug delivery. More interestingly, the MSNs-CDs nanohybrid was able to selectively target folate receptor-overexpressing cancer cells (e.g., HeLa), indicating that folic acid still retained its function even after undergoing the solvothermal reaction. Benefited by these excellent properties, the fluorescent MSNs-CDs nanohybrid can be employed as a fluorescence-guided nanocarrier for the targeted delivery MSNs-CDs@DOX Cancer cell targeted delivery Endocytosis DOX release FL imaging Enhanced chemotherapy FA receptor Cytoplasm N u c le u s pH of anticancer drugs (e.g., doxorubicin), thereby enhancing chemotherapeutic efficacy and reducing side effects. Our studies may provide a facile strategy for the fabrication of multifunctional MSN-based theranostic platforms, which is beneficial in the diagnosis and therapy of cancers in future.