Changes in morphology and organization of collagen fibers contribute to the formation of a microenvironment which facilitate tumor progression through the impact on migration and polarization of the cells [1]. Changes in morphology and organization of collagen fibers in cancer, itself, are the subject of numerous studies, while it is far less known about the changes in collagen fibers in the uninvolved mucosa away from cancer The main histochemical staining for detection of collagen fibers under light micrsocpy is Masson trichrome staining. Recently, second harmonic generation (SHG) imaging of collagen fibers using nonlinear laser scanning micrscopy emerged as a poweful tool enabeling imaging of collagen fibers in unstained and unfixed tissue [2]. Due to growing interest in role of collagen fibers in cancer progression, the number of methods for quantification of different parameters of collagen fibers is increasing. Currently available methods are based on the intensity derivates, intensity variation, Fourier transform, Hough transform, directional filters and fiber tracking algorithm [3]. The aim of our study was to analyse changes in morphology and organization of collagen fibers in the uninvolved colonic mucosa 10 cm and 20 cm away from the cancer, in comparison with healthy subjects, using Masson trichrome staining, SHG imaging and multiple complementary methods for quantification.