A method for calibrating a laser deflection device in a reflected light device of a microscope of a laser microdissection system having a digital image capturing unit comprising an image evaluation module includes generating a laser beam; guiding the laser beam through a microscope objective; directing the laser beam to a position defined by actuation signals; placing a calibration object in the object plane of the microscope objective; actuating the laser deflection device using first actuation signals and first calibration values, making at least one calibration mark on the calibration object; capturing an image of the calibration object by the digital image capturing unit; determining actual position values for the at least one calibration mark: and determining second calibration values based on a relationship between the default position values and the actual position values.