Single-plain video used for measurements of epicardial strains is a technique that yields minor interference with the studied mechanical properties of the ventricle. Due to its low temporal resolution, the existing technique is, however, not appropriate for small animals. We questioned whether the technique could be improved enough to cope with higher heart rates and miniaturization necessary for experiments on rats, mice and guinea pigs. Therefore, we developed a high-speed video system and used it for measuring epicardial strains in guinea pig hearts in situ with the open chest. The improvement was achieved in video hardware (camera: Dalsa D6-0256; framegraber: EPIX PIXCI D32) and software, the markers (glowing acrylate crystals; diameter approximately 0.15 mm) and illumination (UVA light, OSRAM L). Three markers were attached onto the epicardium in the equatorial region of the left ventricular free wall, 1.5 mm apart, with fibrin glue. From their coordinates, we calculated two-dimensional finite strains with end diastole as the reference point. The accuracy of the displacement measurement of the technique and the error introduced by approximate-visual estimation of the left ventricle coordinate system were evaluated. The accuracy of the displacement measurement was +/-1.6 microm and the temporal resolution was 2 ms. Error due to approximate coordinate system orientation was +/-3% of the strain amplitude. The typical amplitude of strains was -0.06, -0.11 and 0.04 in circumferential, axial direction and in-plane shear, respectively. The improvements enable us to perform physiologically relevant measurements of epicardial deformations on guinea pig heart.