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An optical method for measuring the birefringence of muscle fibers was developed, which is realized on an automated Linnick interferometer microscope equipped with a laser. It was shown that the method has some advantages over the methods based on measurements of the intensity of light passing through a crossed polarizer, an analyzer, and a fiber (light polarized microscopy). The method involves direct phase measurements of optical path length at the parallel and perpendicular orientations of the polarization plane of probing radiation. The phase image is reconstructed automatically from interferograms with the use of the four-frame phase-shifting algorithm. The phase images of one and the same central part of the fiber at different orientations of the polarization plane represent two-dimensional numerical maps of the optical path length. The subtraction of these images gives a two-dimensional map of the phase shift, which includes information about the birefringence of the fiber. A formula for birefringence measurements was deduced, which has a certain advantage in comparison to that used earlier in that it does not take into account the thickness of a fiber that depends on the measurement point. The birefringence is normalized to a value of the half sum of phases, which are measured separately in the course of the experiment.