中文摘要：

目的：采用两种骨组织双重染色方法研究斑马鱼颅骨骨组织的发育特征，找出适合于不同发育时期斑马鱼骨组织的染色方法。方法：将不同发育阶段的斑马鱼分为两组，分别用A、B两种方法进行染色，体视显微镜下观察染色结果并拍照。结果：不同发育阶段的斑马鱼分别经A、B染色法染色软骨都被染成蓝色，但两者在骨骼上的染色有差别，对于15d以内骨化不完全的斑马鱼适合用A染色法，15d以上的斑马鱼骨化程度加重更适合用B染色法突出骨骼特征。整体来看，斑马鱼颅骨的骨化过程存在特定的程序化特征，即不同的颅骨组织按照一定的时间顺序逐步完成骨化。结论：可利用颅骨的程序化骨化特征研究人类颅颌面畸形，积极开展口腔医学领域斑马鱼模式生物学的研究。

英文摘要：

Objective： To study the development characteristics of zebrafish calvarial bones in the different developmental stages and compare two double--staining methods. Methods： The zebrafish were collected from 3d to 45d and stained with A and B methods for the bone staining. The staining results were observed with stereomicroscope. Results： Both methods showed no difference in cartilage staining, but bone staining results with B methods was bet- ter in zebrafish older than 15d. The craniofacial bones gradually became calcified in a temporal and spatial order. Conclusion： The temporal and spatial calcification characteristics of craniofacial bone development of zebrafish enable us to study the human craniofacial diseases. Further studies with zebrafish model should be carried out in the basic oral medicine research.