Polarizing microscopy of Picrosirius stained bone sections as a method for analysis of spatial distribution of collagen fibers by optical diffractometry.

Cross sections of femur diaphysis obtained from control and osteopetrotic rats were stained with hematoxylin-eosin (HE) and Picrosirius (SR). Analogous selected areas of bone sections photographed under a polarizing microscope were analysed by optical diffractometry. Since the collagen fibers are a good marker for the structure of bone tissue, their spatial distribution evaluated by optical diffractometry provides information on the tissue architecture. The Picrosirius staining technique enhances the natural birefringency of collagen fibers. Therefore, in the polarizing microscope, pictures of high contrast are obtained. This procedure, by increasing the amount of information in the image, increases the quantity of the data obtained by optical diffractometry in comparison with the HE staining method. The results obtained prove that SR staining combined with polarizing microscopy might be useful for optical diffractometry in analysis of the spatial distribution of collagen fibers in all connective tissues, where they could serve as markers of tissue architecture.