A precise recording of electrophysiological signals requires high-performance flexible bioelectrodes to build a robust skin interface. The past decade has witnessed encouraging progress in the development of elastomeric electrodes for wearable electrophysiological monitoring; however, it remains challenging to achieve excellent flexibility, conformal contact, and high durability simultaneously. Herein, we report on an effective method to fabricate flexible yet durable microneedle electrodes (MEs) based on vertically aligned gold nanowires (Au NWs) embedded polyimide (PI), which meet the above three design requirements. The Au NWs embedded PI MEs could build conformal contact with human skin and maintain electrical stability with minimal contact impedance by effectively penetrating the stratum corneum of the skin. In comparison studies, we found our MEs outperformed conventional gel or elastomeric soft electrodes. We further integrated the vertical Au-NW MEs into a wearable healthcare system and achieved wireless real-time recordings of electromyography (EMG) and electrocardiography (ECG) with high signal-to-noise ratios (SNRs) and low motion artifacts. Our fabrication strategy opens a new route to improve the durability and reliability of emerging nanomaterial-based soft bioelectrodes for long-term wearable healthcare applications.