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目的 探讨低温微波硝酸脱钙方法对骨及软组织抗原活性的影响。方法 根据实验条件分为 4组 :低温微波 10℃组 (Ⅰ组 ) ;冰箱 4℃ (Ⅱ组 ) ;室温 2 0℃ (Ⅲ组 ) ;及温箱 37℃ (Ⅳ组 )。分别使用 5 %硝酸、甲酸 -甲醛混合液、2 0 %甲酸液、Jenkins液、10 %乙烯二胺四乙酸钠 (EDTA)作为脱钙液。应用免疫组织化学LSAB染色和图像分析技术检测ANP ,5 HT ,S 10 0 ,NF ,GFAP和Vimentin的染色结果。结果 以硝酸作为脱钙剂时 ,Ⅰ组 (1.89± 0 97)阳性细胞的光密度值与Ⅱ组 (1 81± 1 0 2 )相近 ,均显著高于Ⅲ (1 0 3± 0 85 )、Ⅳ (0 76± 0 85 )组 ;脱钙液的温度越高 ,阳性细胞的光密度值越低 ,脱钙时间越短。低温微波硝酸与各种脱钙液的染色结果之间无明显差异 ,但脱钙时间显著缩短。结论 低温微波硝酸脱钙能够在较短时间内获得满意的染色效果。
Objective To investigate the effect of low temperature microwave nitric acid decalcification on the activity of bone and soft tissue antigen. The method was divided into four groups according to the experimental conditions: low temperature microwave 10 ℃ group (group Ⅰ), refrigerator 4 ℃ (group Ⅱ), room temperature 20 ℃ (group Ⅲ) and incubator 37 ℃ (group Ⅳ). 5% nitric acid, formic acid-formaldehyde mixture, 20% formic acid solution, Jenkins solution and 10% ethylenediaminetetraacetic acid (EDTA) were used as decalcification solution respectively. Immunohistochemical LSAB staining and image analysis were used to detect the staining results of ANP, 5 HT, S 10 0, NF, GFAP and Vimentin. Results When using nitric acid as decalcification agent, the optical density of positive cells of group Ⅰ (1.89 ± 0.97) was similar to that of group Ⅱ (1 81 ± 102), which was significantly higher than that of group Ⅲ (103 ± 0 85) Ⅳ (0 76 ± 0 85). The higher the decalcification temperature, the lower the optical density of positive cells and the shorter the decalcification time. There was no significant difference between the staining results of low temperature microwave nitric acid and various decalcification solutions, but the decalcification time was significantly shortened. Conclusion Low temperature microwave nitric acid decalcification can obtain satisfactory dyeing effect in a short time.