不同脱钙条件对骨组织免疫组织化学染色抗原性的影响

Antigenicity of Osseous Tissue Influenced by Immunohistochemical Staining under Different Decalcification Conditions

目的:探讨不同脱钙条件对骨组织免疫组织化学染色抗原活性的影响。方法:用8%硝酸分别在冰箱(4℃)、室温(2 0℃)、温箱(37℃)、温箱(6 0℃)、超声波、微波、光波、光波/微波组合Ⅰ组、光波/微波组合Ⅱ组条件下脱钙,应用LSAB免疫组织化学染色技术检测ANP、5 HT、S 1 0 0、NF、GFAP、CD3 1 、CD3 4 和Vimentin的表达,并对其染色结果进行图像分析和统计学处理。结果:8%硝酸脱钙从4℃、2 0℃、37℃到6 0℃随着脱钙液温度的温度升高,脱钙时间逐渐缩短,光波、微波和超声波中脱钙所用的时间比2 0℃、37℃、6 0℃明显缩短,比光波/微波组合Ⅰ、Ⅱ中所用的时间长。光波/微波组合Ⅰ组、Ⅱ组脱钙的免疫组化染色结果均明显比2 0℃、37℃、6 0℃、微波组、光波组和超声波组的好(P <0 .0 1 ) ,而光波/微波组合Ⅰ组和Ⅱ组之间,微波组、光波组和超声波组之间的免疫组化染色结果均无明显差异(P >0 .0 5 )。结论:光波/微波组合Ⅱ 8%硝酸脱钙所用时间最短,免疫组化染色效果最好。

Objective To investigate the antigenicity of osseous tissue influenced by i mmunohistochemical staining method under different decalcification conditions.Methods Decalcifying agent with 8 % nitric acid(NA) was carried under different decalcification conditions: refrigerator(4℃), room temperature(20℃),baker(37℃),baker(60℃),ultrasonic,microwave,lightwave,lightwave/microwaveⅠ and lightwave/microwaveⅡgroup respectively.The expression of ANP,5-HT,S-100,NF,GFAP, CD 31 , CD 34 , and Vimentin was detected byi mmunohistochemical LSAB method and image analysis under different decalcification conditions.Results As the temperatures of decalcifying fluids increased(from 4℃,20℃,37℃,to 60℃) by 8 % NA, the time of decalcification was shortening gradually.The decalcification time with lightwave, microwave,or ultrasonic method was apparently shorter than that under room temperature(20℃),37℃,or 60℃, but was longer than that with lightwave/microwaveⅠor Ⅱ method.The i mmunohistochemical staining effect with lightwave/microwaveⅠor Ⅱ method was significantly better than thatunder room temperature(20℃),37℃, 60℃ or with microwave, lightwave, or ultrasonic method (P<0.01). The i mmunohistochemical staining between lightwave/microwave Ⅰand Ⅱ method or among microwave, lightwave, ultrasonic method was no significant difference(P>0.05).Conclusion With NA as decalcifying agents the decalcification time with lightwave/microwave Ⅱmethod was the shortest and the effect of i mmunohistochemical staining was best among those different decalcification methods.