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This is the first report on a nanofibrous surface patterning process using nano-meshed polymer microcapsules for biological applications. The nano-meshed microcapsules were formed using a method named phase separation assisted electrospray. Features of the nano-meshed microcapsules were found to be tunable by adjusting process conditions. Contrary to non-woven mats formed of continuous nanofibers via conventional electrospinning, nano-meshed microcapsule reported herein enabled a single step nanofibrous surface patterning by electrostatic focusing following the microcapsule formation. The biodegradable polylactic acid made microcapsules were patterned onto a glass substrate with up to 500 mum resolution and human hepatocyte cells were cultured on the patterned areas to confirm biocompatibility of the microcapsule.