For precise X-ray diffraction (XRD) measurement giving the three-dimensional structure of proteins, it is important to prepare high-quality single crystals with suitable shape. As a new processing technique to obtain such protein crystals, we employed femtosecond laser-induced cleaving of protein crystal in a growth vessel containing water solution. An intact protein crystal was precisely processed without mechanical contact in its sealed growth vessel by focusing femtosecond laser pulses. We confirmed that three-dimensional processing of the crystal in its supersaturated solution was realized using multiphoton absorption and that the processing was efficiently enhanced by the cleaving behavior attributed to a photomechanical mechanism of the femtosecond laser ablation. (C) 2007 Elsevier B.V. All rights reserved.