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The authors report a technique for measuring Young’s modulus of a single electrospun nanofiber using the vibrations of two microcantilevers coupled with the nanofiber. The modulus is calculated from the resonant frequency shift resulting from the nanofiber. Polyacrylonitrile nanofibers (200 nm diameter) were collected during electrospinning and wrapped on two similar microcantilevers causing a shift in first resonance from 10.0 to 19.4 kHz. Finite element analysis was used to analyze the frequency shift using images from a scanning electron microscope giving a modulus of the as-spun polyacrylonitrile nanofiber of 26.8 GPa.