The electrical behavior of devices based on highly crystalline thin films of organic semiconductors is inherently anisotropic. Thin film optimization requires simple and accessible means to characterize the orientation of the constituent crystals. The standard polarized light microscopy (PLM) provides a contrast between different crystallites but fails to distinguish crystals with relative orientation of 90°. In this paper, we discuss two methods that enable the unambiguous identification of crystal orientation in thin films of optically anisotropic materials: PLM with a full-wave retardation plate and differential interference contrast (DIC). The latter is standard on most microscopes and delivers images with high contrast and good color balance.As an illustration, we use DIC to extract the optical properties of highly crystalline thin films of three high-performance organic semiconductors: rubrene, 6,13-bis(triisopropylsilylethynyl)pentacene (TIPS-pentacene) and 2,7-dioctyl[1]benzothieno[3,2-b][1]benzothiophene (C8-BTBT). Building on the relation between optical properties and crystal orientation, we demonstrate how DIC characterizes the in-plane crystal orientation of these thin films. This leads to the identification of the fast growth direction of the crystal front.