Skin interstitial fluid (ISF) is a biofluid with information-rich biomarkers for disease diagnosis and prognosis. Microneedle (MN) integration of sampling and instant biomarker readout hold great potential in health status monitoring and point-of-care testing (POCT). The present work describes an attractive MN sensor array for minimally invasive monitoring of ISF microRNA (miRNA) and Cu2+. The MN array is made of methacrylated gelatin (GelMA) and methacrylated hyaluronic acid (MeHA), and a further divisionally encapsulated miRNA and Cu2+ detection system, and is cross-linked through blue-light irradiation. The MN patch displays good mechanical properties that enable withstanding more than 0.4 N per needle, and exhibits a high swelling ratio of 700% that facilitates timely extraction of sufficient ISF for biomarker analysis. For proof-of-concept, it realizes detection of miRNAs and Cu2+ efficiently and quantitatively in an agarose skin and fresh porcine cadaver skin model. Given the good sampling and in situ monitoring ability, the MN array holds great promise for skin ISF-based applications.