We developed a novel strategy for fabricating poly(3,4-ethylenedioxythiophene) (PEDOT) patterns on various substrates, including hydrogels, via sequential solution procedure without multistep chemical etching or lift-off processes. First, PEDOT nanothin films were prepared on a glass substrate by solution phase monomer casting and oxidative polymerization. As a second step, after UV-induced poly(ethylene glycol) (PEG) photolithography at the PEDOT/PEG interface through a photomask, the hydrogel was peeled away from the PEDOT-coated glass substrate to detach the UV-exposed PEDOT region, which left the UV nonexposed PEDOT region intact on the glass substrate, resulting in PEDOT patterns. In a final step, the PEDOT patterns were cleanly transferred from the glass to a flexible hydrogel substrate by a direct-transfer process based on a second round of gelation process. Using this strategy, PEDOT patterns on ITO glass or ITO film were used to successfully fabricate an electrochromic (EC) device that exhibited stable electrochromic switching as a function of applied potential. Furthermore, PEDOT patterns on hydrogel were used to fabricate all organic, flexible microelectrodes with good electrical properties and excellent mechanical flexibility. Importantly, the conductivity of PEDOT patterns on hydrogel (ca. 235 S cm–1) described here is significantly higher than that previously reported (ca. 20–70 S cm–1). This approach can be easily integrated into various technological fabrication steps for the development of next-generation bioelectronics systems.