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Highly‐aligned luminescent electrospun nanofibers were successfully prepared from two binary blends of PFO/PMMA and PF⁺/PMMA. The PFO/PMMA aligned electrospun fibers showed a core/shell structure but the PF⁺/PMMA fibers exhibited periodic aggregate domains in the fibers. The aligned fibers had polarized steady‐state luminescence with a polarized ratio as high as 4, much higher than the non‐woven electrospun fibers or spin‐coated film. Besides, the PF⁺/PMMA aligned electrospun fibers showed an enhanced sensitivity to plasmid DNA. Such aligned electrospun fibers could have potential applications in optoelectronic or sensory devices.