Noorlander Maril L, Melis Paris, Jonker Ard, Van Noorden Cornelis J F
Academic Medical Center, University of Amsterdam, Departments of Cell Biology and Histology, Amsterdam, The Netherlands.
J Histochem Cytochem. 2002 Nov;50(11):1469-74. doi: 10.1177/002215540205001106.
We have developed a quantitative microscopic method to determine changes in the orientation of collagen fibers in the dermis resulting from mechanical stress. The method is based on the use of picrosirius red-stained cryostat sections of piglet skin in which collagen fibers reflect light strongly when epipolarization microscopy is used. Digital images of sections were converted into binary images that were analyzed quantitatively on the basis of the length of the collagen fibers in the plane of the section as a measure for the orientation of the fibers. The length of the fibers was expressed in pixels and the mean length of the 10 longest fibers in the image was taken as the parameter for the orientation of the fibers. To test the procedure in an experimental setting, we used skin after 0 and 30 min of skin stretching. The orientation of the fibers in sections of control skin differed significantly from the orientation of fibers in sections of skin that was stretched mechanically for 30 min [76 +/- 15 (n=5) vs 132 +/- 36 (n=5)]. The method described here is a relatively simple way to determine (changes in) the orientation of individual collagen fibers in connective tissue and can also be applied for analysis of the orientation of any other structural element in tissues so long as a representative binary image can be created.
我们开发了一种定量显微镜方法，以确定机械应力导致的真皮中胶原纤维取向的变化。该方法基于使用仔猪皮肤经苦味酸天狼星红染色的低温恒温器切片，在使用偏光显微镜时，其中的胶原纤维会强烈反射光线。切片的数字图像被转换为二值图像，并根据切片平面中胶原纤维的长度进行定量分析，以此作为纤维取向的度量。纤维长度以像素表示，并将图像中10条最长纤维的平均长度作为纤维取向的参数。为了在实验环境中测试该程序，我们使用了拉伸0分钟和30分钟后的皮肤。对照皮肤切片中纤维的取向与机械拉伸30分钟的皮肤切片中纤维的取向有显著差异[76±15（n = 5）对132±36（n = 5）]。这里描述的方法是确定结缔组织中单个胶原纤维取向（变化）的一种相对简单的方法，并且只要能创建代表性的二值图像，也可用于分析组织中任何其他结构元素的取向。
