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High diffraction efficiency refractive index-modulated holographic micrograting on a polymer thin film was obtained with a single femtosecond laser pulse interference and photo-polymerization. The diffractive efficiency of the micrograting with an He-Ne laser is up to 78% when the thickness of the film is 90 μm, which corresponds to the refractive index-modulation as large as 2.4 × 10-3. The compared experiment with a femtosecond oscillator laser indicated that a single femtosecond Iaser pulse has the advantages in laser machining over the cw holography method such as high fabrication speed, good surface structure and high diffractive efficiency.