Nonlinear optical microscopy is a new and rapidly growing technique within which ultrafast laser technology finds a wide range of applications. Pulse widening, due to the microscope optics, is an issue of major concern for nonlinear excitation efficiency. We herewith describe a novel, simple and inexpensive autocorrelation technique to characterize the laser temporal behavior at the microscope focal plane. The method is based on a wavefront-division lateral shearing interferometer which is inserted into the microscope optical path like an ordinary filter, while a spatially uniform fluorescent specimen is observed. The two-photon excited fluorescent image provides the second-order autocorrelation curve.