NaOH消蚀/SEM技术在胶原纤维与组织钙化研究中的应用

NaOH-maceration/SEM technique for studies of the relationship between collagen fibers and tissue calcification

应用NaOH消蚀 叔丁醇冻结干燥 SEM法 ,观察了人松果体的钙化及其与胶原纤维的关系。将固定的标本在 2NNaOH溶液 (2 0℃ )中浸泡 5～ 7天 ,蒸馏水浸洗去除细胞成分和结缔组织基质 ,完好地保留胶原纤维和钙化的自然状态 ,然后用叔丁醇冻结干燥和SEM观察。结果清晰地揭示了人松果体内胶原纤维支架和钙化的三维形态以及二者的密切关系。结果提示 ,通过NaOH消蚀 SEM法所获得的胶原纤维与钙化的三维图像 ,较通过光镜和透射电镜方法所得到的二维图像立体感强 ;也较通过冷冻割断 /SEM法所获得的图像清晰。该方法对标本的固定要求不高。叔丁醇冻结干燥法与CO2 临界点干燥法相比 ,更简便、易行。因此 。

We investigated calcification in human pineal body and its relationship to collagen fibers by NaOH maceration/t butyl alcohol freeze drying/SEM method. After immersion of fixed tissues in 2N NaOH(20℃)for 5～7 days and followed by rinsing in distilled water the cellular elements and the ground substance of the connective tissue were removed, exposing collagen fibers and calcification in their natural location. The results specimens were dried by t butyl alcohol freeze drying method. SEM observation of the preparations clearly demonstrated three dimensional architecture of collagen fibers and calcification. The data presented here suggest that the three dimensional images of the collagen fibers and calcification by this method is superior to the two dimensional images by LM or TEM and the three dimensional images by frozen fracture/SEM. Its fixation is simple. The t butyl alcohol freeze drying method is much simpler and easier than the critical point drying method. Therefore, this method could be widely used to study the relationship between collagen fibers and tissue calcification.