In this work we present non-resonant four-wave mixing microscopy as an additional contrast mechanism in nonlinear microscopy. The setup for this technique was based on a commercially available multiphoton microscope setup equipped with a titanium:sapphire-laser and an optical parametric oscillator as light sources. Fundamental system characteristics with respect to the spatio-temporal pulse overlap and the influence of aberrations on the process are presented. Experiments regarding the directionality of the four-wave mixing signal performed on fresh porcine meat showed an average ratio of the backward to forward signal mean intensity of 0.16 ± 0.01. Nevertheless, structural information is comparable for both detection modalities. This highlights the potential of four-wave mixing microscopy for in vivo applications. Furthermore, results on porcine meat show the additional contrast generated by four-wave mixing. In summary, the results show a great potential of non-resonant four-wave mixing microscopy as label-free imaging modality in the biomedical sciences.