Precise nano- and microscale control of the architecture of biodegradable biomaterials is desirable for several biotechnological applications such as drug delivery, diagnostics, and medical imaging. Herein, we combine electrohydrodynamic co-jetting and highly specific surface modification (via Huisgen 1,3-dipolar cycloaddition) to prepare particles and fibers with spatioselective surface modification. We first prepared biphasic particles and fibers from commercial poly(lactide-co-glycolide) copolymers via electrohydrodynamic co-jetting of two organic solutions loaded with fluorescent macromolecules and acetylene-modified PLGA derivatives. (i) Spatially controlled reaction of poly [lactide-co-(propargyl glycolide)] with O-(2-aminoethyl)-O'-(2-azidoethyl)heptaethylene glycol and (ii) subsequent conversion of the newly introduced amino groups with fluorescence probes resulted in particles and fibers with surface modification of one hemisphere only.